专利名称：为抗生素的生物合成从蓝灰链霉菌非产蓝亚种中克隆基因及其使用方法
背景技术：
发明领域本发明涉及编码用于生产LL-F28249化合物的蛋白质的新的生物合成基因及其在从蓝灰链霉菌非产蓝亚种(Streptomyces cyaneogriseus subsp.noncyanogenus)发酵产物中制备活性代谢物中的应用。本发明进一步涉及制备天然代谢物的活性半合成衍生物的生物合成途径的遗传操纵。
背景技术：
本发明所引用的所有的专利和出版物此处均完整引用作为参考。
链霉菌属是大量的具有商业重要性的次级代谢物的生产者，所述次级代谢物包括大量的活性抗生素，如β-内酰胺以及大环内酯化合物或大环内酯物。由于链霉菌产生的次级代谢物的商业重要性，最近已有大量的投资致力于发展对链霉菌的分子遗传操纵的方法。生产过程已经发展为通过由聚乙二醇介导的转化以及通过自大肠杆菌的接合转移导入遗传物质。载体发展成为包括高拷贝和低拷贝数的载体、整合载体以及大肠杆菌-链霉菌穿梭载体。这些链霉菌的分子遗传操纵方法都已经在D.A.Hopwood等所著的链霉菌的遗传操作实验手册，John Innes Foundation Press，Norwich，UK(1985)中作了总结。在许多情况下，链霉菌中产生次级代谢物的基因都是聚集在一起的。因此，在生物合成基因簇中识别单个基因也就容易导致识别全部的负责代谢物生物合成的基因。这一发现被证明具有巨大的价值，次级代谢物的生物合成基因簇已经通过反向遗传学、阻断突变株的互补以及异源探针的抗性和使用被克隆出来。通过这些方法，已获得许多大环内酯物生物合成基因簇的核苷酸以及预测的氨基酸序列数据，包括那些指导红霉素(参见S.Donadio等，Science252675-679(1991)以及S.F.Haydock等，Molecular and General Genetics230120-128(1991))；雷帕霉素(参见T.Schwecke等，Proceedings of the National Academy of Sciences USA927839-7843(1995)以及X.YUAN等，GENE2031-9(1997))；FK506(H.Motamedi和A.Shafiee，European Journal of Biochemistry256528-534(1998))；竹桃霉素(D.G.Swan等，Molecular and General Genetics242358-362(1994))以及利福霉素(参见P.R August等，Chemistry & Biology569-79(1998))合成的基因。然而，编码被称为LL-F28249的大环内酯化合物的完整的生物合成基因簇还没有在现有技术中有所描述。
有许多报道称分子遗传操纵能用于改变聚酮化合物(polyketide)的生物合成进程(参见S.Donadio等，Science 252675-679(1991)和S.donadio等，Proceedingsof the National Academy of Sciences USA 907119(1993))。在这些研究中，红霉素相关的内酯都是在操纵6-脱氧红霉内酯B合酶(“DEBS”)的基因簇(负责红霉素生物合成的核心聚酮合成酶基因簇)后生产的，以至于组件(module)4烯酰还原酶，或是具有更重要的所述作用的组件5酮还原酶的酶域二者其中之一没有功能。含有这些DEBS基因簇突变体的菌株产生所预期的红霉素相关内酯。这些开创性研究自此被重复进行和扩展，许多这些研究的结果在文献中被加以评述(参见，例如，L.Katz和S.Donadio，Annual Reviews of Microbiology47875-912(1993)；C.R.Hutchinson和I.Fujii，Annual Reviews of Microbiology49201-238(1995)；D.A.Hopwood，Chemical Reviews972465-2497(1997)；和C.W.Carreras和D.V.Santi，Current Opinions in Biotechnology9403-411(1998))。
这些文献中总结的数据表明在I型聚酮合成酶(“PKS”)组件中的催化域的排列是保守的，许多高度保守的氨基酸序列基元已经在那些生物合成基因簇中描述过了。例如，除虫链霉菌(S.avermitilis)产生的除虫菌素的生物合成基因簇的排列已经被报道(参见D.J.MacNeil等，Gene.115119-125(1992)以及D.J.MacNeil等，Annals of the New York Academy of Sciences721123-132(1994))，并且所述生物合成基因簇的部分核苷酸序列已经被报道或者是可获得的。MacNeil和他的同事已经预测了这种组件的组织排列并报导了一种野生型蓝灰链霉菌(S.cyaneogriseus(NRRL 15773)nemadectin生物合成基因簇的有限的限制性核酸内切酶图谱(参见D.J.MacNeil等，Annals of the New YorkAcademy of Sciences721123-132(1994))，但是他们的限制性图谱是不完整的。他们的分析仅仅表明存在9个PKS功能的组件重复，以及限定蓝灰链霉菌的基因组的75kb区域所需的6个相互重叠的克隆。MacNeil等没有完成整个生物合成基因簇的DNA序列。作为替换，那些作者们仅仅对所选的粘粒末端进行了测序。从这些有限的序列信息中，他们只能够得出一个非常粗略的限制性酶切图谱。进行了进一步的C-13标记研究，接着提出了一种从其组成的酰基单元中合成LL-F28249α化合物的机制假说(H.R.Tsou等，Journal of Antibiotics(TOKYO)42398-406(1989))。
这种高活性LL-F28249α化合物是从蓝灰链霉菌非产蓝亚种(此后均称之为“蓝灰链霉菌(S.cyaneogriseus”)的发酵肉汤培养基中分离出来的。该化合物是广泛用于治疗线虫以及节肢动物寄生虫的治疗的天然的endectocidal剂，包括控制或防治蠕虫、节肢动物外寄生以及螨虫的感染。由蓝灰链霉菌生产的一系列抗寄生LL-F28249α化合物在结构上与表征清楚的除虫菌素近似，但又与之有明显区别。美国专利5,106,994以及其系列申请美国专利5,169,956描述了主要和次要成分，LL-F28249α-λ，的制备。LL-F28249族化合物进一步包括，但是不限于，在美国专利4916154中出现的半合成23-氧衍生物以及LL-F28249α-λ的23-亚氨衍生物。莫西菌素，化学名称为23-(O-甲基肟)-LL-F28249α，是一种特别有效的23-亚氨衍生物。LL-F28249衍生物的其它例子包括，但不限于，23-(O-甲基肟)-5-(苯氧基乙酰氧基)-LL-F28249α，23-(缩氨基脲)-LL-F28249α和23-(缩氨基硫脲)-LL-F28249α。
作为nemadectin代谢物中主要的一种，LL-F28249α(此后称之为“Fα”)，通过一个四步的化学过程转化为具有商业重要性的化合物莫西菌素(moxidectin)。迄今仍未知的Fα的生物合成基因簇的确定将具有巨大的商业重要性。基因的分离不仅有利于生产活性Fα化合物以及其它LL-F28249族化合物的天然成员，而且可用于更迅速更有效地制备具有商业有效性的半合成衍生物例如莫西菌素。
因此本发明的一个重要的目的就是分离并且识别编码负责产生LL-F28249化合物，尤其是LL-F28249α代谢物的蛋白质的完整的核苷酸序列，然后分离并确定包括生物合成蛋白质的氨基酸序列的功能。
另一个目的是提供一种用于直接从蓝灰链霉菌非产蓝亚种的生物工程菌株的发酵肉汤培养基中分离天然和半合成衍生物的新的途径。
进一步的目的是提供一种比迄今采用的步骤更少的有效的方法来制备莫西菌素。
本发明更进一步的目的和目标呈现在说明书下面的内容中。
前述目的是通过提供一种新的，经纯化分离的核酸分子来实现的，该核酸分子编码产生LL-F28249化合物的完整的生物合成途径相关的蛋白质。
发明简述本发明涉及形成LL-F28249化合物以及(最重要的)高活性的主要成分LL-F28249α的完整生物合成途径的独特(unique)克隆及其表征。完整的DNA基因簇以及其在一个合适的宿主内的表达使得高活性的天然代谢物以及半合成衍生物的有效生产得以实现。尤为显著的是，整个生物合成途径是有效的包含在仅仅三个质粒中，这三个质粒是粘粒11，36和40(此后分别称之为“Cos11”，“Cos36”，“Cos40”)。
附图简介参考相应的附图，本发明的背景及其与现有技术的差别将会在下文加以进一步描述
图1阐明了通过包含在根据本发明所述方法制备的粘粒中的基因片段制备LL-F28249化合物的生物合成基因簇的构建。蓝灰链霉菌粘粒文库是通过连接蓝灰链霉菌基因组DNA的Sal3A片段至粘粒载体pSuperCos1的BamH1位点而构建的。随后产生的粘粒文库被转化到大肠杆菌VCS257中。不同的粘粒通过采用除虫菌素酮脂酰合成酶探针，或是这里所描述的“步行”技术的杂交技术来识别。这些粘粒通过限制性酶切图谱以及DNA测序表征。Fα基因簇的BamH1限制性图谱通过分析重叠的粘粒而获得，又通过DNA测序加以证实。B表示一个BamH1位点。
图2阐明了生物合成蛋白质及其制备LL-F28249化合物的克隆的生物合成基因簇所编码的位点。一大约长88Kbp，含有整个Fα聚酮合成酶基因簇的临近的核苷酸序列是通过对重叠的粘粒及其亚克隆进行测序而得到的。13个组件以及各个酶域通过使用BLAST序列对比分析来鉴定。其它的生物合成基因以同样的方式鉴定。以下缩写在图中使用ACP，酰基载体蛋白；DH，脱水酶；ER，烯酰还原酶；KR，酮还原酶；KS，酮脂酰合成酶；LD，载入域；TE，硫酯酶；MT，甲基转移酶；AT，酰基转移酶。
图3显示命名为pKR0.9的载体的组成结构，其为一在pSL301(Invitrogen，Carlsbad，CA)的BstEII-AatII位点存在一pNE57(含有Fα组件3酮还原酶域的目的区域)的长度900bp的BstEll-AatII片段。以下缩写在图中使用mod3 KR，Fα组件3酮还原酶域；amp，氨苄青霉素抗性标记。
图4显示了pFDmod3/5.2系列的质粒组成结构。这些质粒被构建用于将Fα组件3酮还原酶域的定点突变与侧翼DNA结合起来以促进同源整合。骨架载体是大肠杆菌-链霉素穿梭载体pKC1132。以下缩写在图中使用mod3 KS，组件3酮酯酰合成酶域；mod3 AT，组件3酰基转移酶；mod3 KR，组件3酮还原酶域；apra，阿泊拉霉素抗性标记。
图5显示了pFDmod3/4.2系列的质粒组成结构。这些质粒通过将pFDmod3/4.2系列质粒移除长度大约为1Kbp的侧翼DNA以减少异常的整合而衍生得到。以下缩写在图中使用mod3 AT，组件3酰基转移酶；mod3 DH，组件3脱水酶；mod 3 ER，组件3烯酰还原酶；mod3 KR，组件3酮还原酶域；apra，阿泊拉霉素抗性标记。
图6表明了用于制备LL-F28249化合物的生物合成基因的全长核苷酸序列(88400bp)(相应于序列SEQ ID NO1)。
图7表明了由ORF1基因(对应于序列SEQ ID NO2)编码的调节蛋白质的推定氨基酸序列(922aa)。
图8表明了由ORF2基因(对应于序列SEQ ID NO3)编码的硫酯酶蛋白质的推定氨基酸序列(259aa)。
图9表明了由ORF3基因(对应于序列SEQ ID NO4)编码的还原酶蛋白的推定氨基酸序列(267aa)。
图10表明了由ORF4基因(对应于序列SEQ ID NO5)编码的Mod1的载入域蛋白质的推定氨基酸序列(2341aa)。
图11表明了由ORF5基因(对应于序列SEQ ID NO6)编码的Mod2-Mod3的载入域蛋白质的推定氨基酸序列(3723aa)。
图12表明了由ORF6基因(对应于序列SEQ ID NO7)编码的Mod4-Mod7的载入域蛋白质的推定氨基酸序列(6043aa)。
图13表明了由ORF7基因(对应于序列SEQ ID NO8)编码的甲基转移酶蛋白质的推定氨基酸序列(284aa)。
图14表明了由ORF8基因(对应于序列SEQ ID NO9)编码的p450蛋白质的推定氨基酸序列(468aa)。
图15表明了由ORF9基因(对应于序列SEQ ID NO10)编码的Mod8-Mod10的载入域蛋白质的推定氨基酸序列(5674aa)。
图16表明了由ORF10基因(对应于序列SEQ ID NO11)编码的Mod11-Mod13的载入域蛋白质的推定氨基酸序列(5166aa)。
图17表明了由ORF11基因(对应于序列SEQ ID NO12)编码的氧化还原酶蛋白质的推定氨基酸序列(254aa)。
发明详述按照本发明，提供了一种新的，经过纯化、分离的核酸分子，其编码用于产生LL-F28249化合物的完全生物合成途径中的蛋白质。本发明的核酸分子是从产生抗生素的野生型或突变型链霉菌中分离出来的。令人惊奇的是，用于编码所有必需生物合成蛋白质的完整的DNA仅在三个粘粒中得以有效地包装。已构建的含有本发明所述核酸分子的这三个粘粒，Cos11、Cos36和Cos40足以重新建立整个用以产生LL-F28249化合物的途径。因此本发明独特的提供了在三个粘粒中的完整的生物合成基因簇，如一优选实施方案所提到的，该完整的生物合成基因簇使得一种比以前所构思的步骤更少、实质上更加有效的制备活性抗寄生LL-F28249化合物，尤其是莫西菌素，的方法成为可能。本发明的成功克服了现有技术中其他人试图分离完整生物合成基因的失败的努力，满足了长期存在的需求。
该完整的DNA基因簇的核苷酸序列(对应于序列SEQ ID NO1)已经在附图6中进行了充分的描述。本发明的范围还包括其互补链，也就是那些互补的核苷酸序列(例如，A取代T，C取代G，反之亦然)和/或反义核苷酸序列(例如，一个降序排列而不是正向或是升序链，举个例子，改变阅读方向由5′到3′变为由3′到5′)。
本发明进一步包括与从微生物源分离的权利要求4的核酸序列杂交，并且编码产生LL-F28249化合物的生物合成途径的蛋白质的核酸分子。本领域普通技术人员公知的典型的杂交步骤和条件在Sambrook等，分子克隆实验手册，第二版，Cold SpringHarborLaboratoryPress，Cold SpringHarbor，NY(1989)中进行了描述。同源探针采用的是标准或严谨杂交条件，而与靶核苷酸序列的同源性低于100％的部分同源探针则采用非严谨杂交条件。在后者的部分同源探针实施方案中，一系列Southern和Northern杂交可以很容易的在不同严谨程度条件下进行。例如，当在含有甲酰胺的溶液中杂交时，优选的条件采用温度和离子浓度为恒定的大约42℃且溶液含有6X SSC，甲酰胺浓度为50％。非严谨杂交条件可以采用同样的温度和离子浓度，但是在退火缓冲液中使用更少或更低量的甲酰胺，其浓度在大约45％至0％的范围。可选的，杂交可以在不含甲酰胺的水溶液中进行。通常水溶液中的杂交，溶液中的离子浓度保持恒定，通常是大约1M Na+，而退火温度可以从大约68℃降低至42℃。
一般而言，基因组DNA的分离和表征以及来源于合适宿主的克隆的重组DNA可以通过标准的或严谨杂交技术获得。所述标准的严谨杂交技术使用全部或者部分核苷酸序列作为探针筛选合适的文库。作为一种两者择一的办法，基于其它相关已知的DNA和蛋白质序列构建的寡核苷酸引物可以被用于聚合酶链反应以扩增和识别其它相同或相关序列。这里所述的核苷酸和蛋白质是通过常规方法分离和纯化，从而得到不同纯度的产物。优选的，获得蛋白质的实质纯形式，但是大约80％-90％的较低的纯度也是可以接受的。本发明的范围还包括由化学合成制得的DNA和蛋白质，这些DNA和蛋白质与从产生抗生素的野生型或突变链霉菌直接衍生的且经过常规检测或者标准测定证实的那些物质具有相同或实质上相同的结构，所述常规检测或者标准测定是包含在LL-F28249化合物生物合成途径中的。
此外，本发明包括以及完整的描述了分离的包括氨基酸序列的生物合成蛋白质。所述蛋白质包括，但不局限于由ORF1基因(序列对应于SEQ ID NO2)编码的调节蛋白，由ORF2基因(序列对应于SEQ ID NO3)编码的硫酯酶蛋白，由ORF3基因(序列对应于SEQ ID NO4)编码的还原酶蛋白，由ORF4基因(序列对应于SEQ ID NO5)编码的Mod1的载入域蛋白，由ORF5基因(序列对应于SEQ ID NO6)编码的Mod2-Mod3的载入域蛋白，由ORF6基因(序列对应于SEQ ID NO7)编码的Mod4-Mod7的载入域蛋白，由ORF7基因(序列对应于SEQ ID NO8)编码的甲基转移酶蛋白，由ORF8基因(序列对应于SEQ ID NO9)编码的p450蛋白，由ORF9基因(序列对应于SEQ IDNO10)编码的Mod8-Mod10的载入域蛋白，由ORF10基因(序列对应于SEQID NO11)编码的Mod11-Mod13的载入域蛋白，以及由ORF11基因(序列对应于SEQ ID NO12)编码的氧化还原酶蛋白。
编码生物合成蛋白的染色体组DNA簇的开放阅读框，可以通过大量的本领域所公知的技术来识别。这些技术包括，但不局限于定位已知起始和终止密码子的计算机分析，基于密码子频率推定的阅读框定位，以及与其它已知链霉菌株中表达的基因的相似性序列比较等等。照这样，本发明的蛋白质用本发明的核苷酸序列识别，然后可以通过分离和纯化以及可选的，通过化学途径合成得到开放阅读框或编码的蛋白质。基于开放性阅读框以及合适的启动子，起始密码子，终止子等等的可表达的基因构建体可以被设计并被导入合适的宿主从而表达由开放阅读框编码的蛋白质。
这里所述的术语“蛋白质”是指多肽，酶等等，如本领域通常使用的这些术语的含义。它们由包括产生LLF28249化合物的生物合成途径的核酸分子所编码。本发明的蛋白质包含不同长度的氨基酸链，其中包括全长序列，所述的氨基酸残基通过共价肽键连接，也包括其生物活性变异体。蛋白质可以是天然的，重组的或是合成的。例如，生物合成蛋白可以通过在合适的表达载体中插入编码蛋白质的核酸序列并且在适当的宿主中表达蛋白质的常规的重组技术，或是通过标准化学合成方法，即在Merrifield，J.Am.Chem.Soc.852149-2154(1963)中描述的Merrifield的固相合成法来制备，其中氨基酸是逐个依次连接到一条氨基酸链上的。可选的，为了实现自动合成蛋白质，可从各制造商，如Perkin-Elmer，Inc.(Wellesley，MA)购得现代设备。
那些包括在本发明范围内的生物活性变异体包括，至少包括，由本发明的核酸分子编码的氨基酸序列的生物功能部分。这里所述的“生物功能部分”是指仍然保留了蛋白质活性功能的蛋白质结构部分，举个例子，由ORF1基因编码的调节蛋白质分子的部分，具有相同或基本上相同的活性和/或结合特性的，也就是，至少大约90，并且更优选的95％的相似性或效能。蛋白的生物活性变异体包括经过缺失、替换或增加氨基酸残基的活性氨基酸结构以及天然的等位基因等等。可以通过对全长蛋白质进行化学或酶消化制备片段，然后以标准测定法检测那些片段，从而分析氨基酸结构的哪一部分保留有与全长蛋白质相同或基本上相同的生物活性，藉此可以很容易的确定生物功能部分。
迄今未知的Fα的完整的生物合成基因簇的鉴定具有巨大的商业重要性。依照目前的方法，对基因的分离以及详尽描述提高了活性Fα化合物和化合物LL-F28249家族的其它天然成员的产量。此外，基因的信息使得以比以往化学生产过程更为快速而有效的方式制备具有商业有效性的半合成衍生物例如莫西菌素的改良的方法成为可能。在此描述的，作为克隆和表征新的Fα生物合成基因簇的直接而有益的结果，可以获得采用蓝灰链霉菌的生物工程菌株得到莫西菌素的直接发酵产物和其它LL-F28249衍生物的独特的方法本发明的一个优势在于提高蓝灰链霉菌发酵肉汤产生的高活性的Fα的产量。Cos11含有一个推定的PKS簇的转录活化基因(ORF 1)。提高活化因子的表达水平会导致Fα产量的提高。这可以通过已知技术(参见，例如Perez-Llarena等，Journal ofBacteriology1792053-2059(1997))提高基因的拷贝数或是增强该基因的调节序列元件来实现。
另一个由本发明的全长生物合成基因簇带来的有益效果是使得天然和半合成的LL-F28249族化合物的衍生物的高效发酵产量以及生产成为可能。天然和半合成的LL-F28249族化合物的衍生物，例如LL-F28249α，LL-F28249β，LL-F28249γ，23-(O-甲基肟)-LL-F28249α(莫西菌素)，23-(O-甲基肟)-5-苯氧基乙酰氧基)-LL-F28249α，23-(缩氨基脲)-LL-F28249α，23-(缩氨基硫脲)-LL-F28249α等等。在编码负责生产LL-F28249化合物的蛋白质以及作为主要产物的理想的Fα代谢物的生物合成基因的识别过程中，对途径的额外的克隆和诱变容易产生作为发酵过程的副产物的其它代谢物。生物合成基因对于减少制备所述族中其它半合成成员的化学反应的步骤是特别有用的。
本发明特别优选的用途包括，采用比以往所知的化学途径更少的步骤制备具有商业重要性的化合物莫西菌素。莫西菌素现在由Fα通过四步化学步骤生产出来，Fα首先是通过发酵蓝灰链霉菌非产蓝亚种制得。由天然代谢物Fα向莫西菌素的转变过程包括以下化学反应(1)5-羟基的保护；(2)23-羟基氧化成一个酮官能团；(3)转化23-酮为23-O-甲基肟以及(4)5-羟基的解保护。本发明的有效方法现在允许23-酮Fα向莫西菌素的化学转化在一个步骤中完成。
通过产生生物合成基因簇突变体，降解LL-F28249化合物结构中位点23的酮官能团的特定活性被除去，并且化学合成也减少到一步。令人惊奇的是，聚酮合成酶的组件的剩余物保留了功能并且保留的聚酮合成酶功能识别非天然聚酮化合物中间体。可以使用、克隆以及再次使用这种独特的生物工程菌株，从而获得23-酮Fα的直接发酵产物，进一步减少正常的程序时间。
在下面的例子中，所选择的诱变阐明了如何修饰Fα生物合成以及如何通过现有方法获得所需代谢物。基本上，蓝灰链霉菌Fα生物合成基因簇的组件3酮还原酶域的突变体是通过定点位点诱变获得的。这些酮还原酶变异体是通过将预测的Fα的组件3酮还原酶域与来自具有生物活性的酮还原酶域以及几个“隐藏的”酮还原酶域的比较而设计出来的。然后通过同源重组，将蓝灰链霉菌Fα生物合成基因簇的组件3酮还原酶域替换成那些变异域，从而改变Fα的生物合成以及获得所需代谢物。
一般而言，定点位点诱变会在23-酮(氧)还原酶基因(23-KR基因)引入一个小的缺失或是点突变，这导致23-酮还原酶域没有功能，却保留了聚酮合成酶的其它域的功能。通过标准的方法将23-KR基因突变引入蓝灰链霉菌非产蓝亚种野生型菌株或是Fα生产菌株142的突变体中，从而直接发酵获得23-酮(氧)Fα产物。此外，携带23-KR基因突变的整个FαPKS基因簇可以被导入到合适的宿主细胞，例如浅青紫链霉菌(S.Lividans)，天蓝色链霉菌(S.coelicolor)，大肠杆菌等等以生产23-酮Fα。为了进一步得到23-酮Fα的发酵产物，转化的宿主细胞被用作通过所述方法接合转移蓝灰链霉菌的DNA的来源。
23-含氧化合物的亚氨基衍生物(23-肟)很容易通过标准技术制备，这些技术如M.McElvain The Characterization of Organic Compounds，published byMacMillian Company，New York，1953，pages 204-205中有描述此处完整引用作为参考。通常情况下，在室温到大约50℃的条件下，23-含氧化合物在存在乙酸以及过量的氨基衍生剂的情况下，与等量的乙酸钠混合，搅拌溶于醇或二氧六环。所述醇为甲醇或乙醇；所述氨基衍生剂为，例如盐酸羟胺，O-甲基盐酸羟胺，盐酸氨基脲等等。在室温下，反应通常需几小时至几天完成，但是可以容易的通过加热来加速。令人惊奇的是，接下来的经由23-酮Fα化合物向莫西菌素的转化是唯一一个必需的化学反应。
进一步考虑，通过本领域技术人员公知的遗传操作方法，将三个粘粒Cos11，Cos36和Cos40中含有的遗传物质整合到两个或一个质粒。举个例子，通过本发明的方法制备的三个粘粒Cos11，Cos36和Cos40中克隆的Fα生物合成基因能够在两个或是一个质粒中装配成完整的聚酮合成酶(PKS)基因簇。该装配可以通过使用克隆，PCR或是合成基因或其它任何本领域所熟知的技术的组合来获得。装配的Fα PKS基因簇可以被导入到合适的宿主以生产Fα，合适的宿主例如浅青紫链霉菌，天蓝色链霉菌，大肠杆菌等等。此后，装配的PKS基因簇可以用于无细胞表达系统以生产更多的Fα以及相关产物，例如Olsthoom-Tieleman等在Eur.J.Biochem.2683807-3815(2001)中描述的一种无细胞表达系统。
通过使用核心LL-F28249α聚酮合成酶的组件结构以及这些组件内的功能域，这里所述的生物合成基因簇得以克隆和充分的表征。一般说来，为了分离生物合成基因，采用商业化的载体pSuperCos(Stratagene，La Jolla，CA)构建蓝灰链霉菌的基因组DNA粘粒库。该粘粒库与除虫菌素组件1酮酯酰合成酶相应的DNA片段制备的探针进行杂交，除虫菌素组件1酮酯酰合成酶是通过聚合酶链反应从除虫链霉菌基因组DNA中扩增出来的。随后，使用聚合酶链反应，从前述表征的粘粒中扩增的Fα生物合成基因簇的几个区域被用作探针，以分离其它的粘粒。通过使用这些方法，一系列叠加起来跨越超过100Kbp的基因组DNA的粘粒被分离出来。通过完整的限制性酶切图谱以及彻底的核苷酸序列分析，对粘粒进行鉴定，给出了精确而又明确的跨越长度接近88Kbp的毗邻核苷酸序列。对该核苷酸序列的分析表明，组件聚酮合成酶的13个完整组件与其它至少6个参予Fα的生物合成或其调节的基因在一起。
本发明进一步包括含有本发明核酸的生物功能质粒或载体。本发明的特定质粒是根据它们整合大DNA基因簇的能力筛选出来的，但是它们是常见的质粒，并且是从一般的载体如pKR0.9，pFDmod3/5.2系列，pFDmod3/4.2系列等等衍生而来的。
尽管在实施例中大肠杆菌被用作异源宿主，但抗生素生物合成基因的异源表达可以在广泛的放线菌目，杆状菌，棒杆菌，嗜热放线菌属等等中发生，只要它们能够容纳转化的所述的大质粒构建体。那些被转化的宿主包括，但不局限于青紫链霉菌，天蓝色链霉菌，灰褐链霉菌(Streptomyces griseofuscus)以及产二素链霉菌(Streptomyces ambofaciens)，这些都是已知相对来说没有限制的。优选的是，能被质粒或载体稳定转化或转染的合适的宿主细胞是天蓝色链霉菌或大肠杆菌-链霉菌粘粒载体。蛋白质的体外表达可以采用标准的工艺方法。
以下的部分关注于对于本领域普通技术人员可获得的常规方法以及材料，这些方法和材料已经被成功地用于克隆和表征本发明所述的完整的、大的生物合成途径。
一般的方法和材料A材料，质粒以及菌株一种含有在大肠杆菌和链霉菌中复制和筛选所需的因子的大肠杆菌-链霉菌穿梭载体pKC1132被用于本实验(参见M.Bierman等，Gene11643-49(1992))的整个过程中，该穿梭载体包含用阿泊拉霉素筛选的抗生素抗性标记，。除了pKC1132之外，商业化的克隆载体可以采用，如上所述应用。本领域普通技术人员可以选用其它已知的载体(可以很容易的被实施例中的载体替换)，避免或缩小标准方法所使用的旧的粘粒锚定大肠杆菌菌株中存在的不稳定问题。采用与在其它质粒上建立的方法类似的方法来对质粒进行操作。典型的步骤记载在Sambrook等，分子克隆实验手册，第二版，Cold Spring Harbor LaboratoryPress，Cold Spring Harbor，NY(1989)中。链霉菌属的典型的步骤记载在D.A.Hopwood等，链霉菌的遗传操作实验手册，John Innes Foundation Press，Norwich，UK(1985)中。除非与前述参考的实验手册相同，本项工作中使用的特殊的方法这里会做描述。
贯穿于本项工作中使用的常见的实验室菌株大肠杆菌JM109和DH5α很容易从许多商业途径(例如Stratagene，La Jolla，CA)获得。大肠杆菌XL1-BlueMRF′株从Stratagene(La Jolla，CA)获得。大肠杆菌ETS12567(pUZ8002)是从华盛顿大学(Seattle，WA)化学系的Heinz Floss教授处获得。大肠杆菌VCS257是从Stratagene(La Jolla，CA)获得。除虫链霉菌来源于美国典型培养物保藏中心，保藏号为No.31,267；但也可以由农业研究培养物保藏中心(NRRL)，1815N.University street，Peoria，IL61604获得，其编号为8165。“野生型”蓝灰链霉菌非产蓝亚种LL-F28249(NRRL 15773)以及命名为“蓝灰链霉菌菌株142”的蓝灰链霉菌的突变Fα生产菌在整个本发明的公开的内容内都是分别使用的但是它们是可以互换的，并且可以在所公开的任意一个特定步骤中互相替换。衍生自野生菌株的菌株142采用典型的遗传操作以提高抗生素产量，但是它仍保留了与野生菌株相同的聚酮合成酶DNA序列。由于它们的聚酮合成酶序列相同，因此所有这里所述质粒，包括，但不局限于Cos11，Cos36 and Cos40可以衍生自野生型蓝灰链霉菌非产蓝亚种或是蓝灰链霉菌株142，并获得相同的结果。
B.质粒DNA的限制性分析质粒DNA的限制性分析的步骤，琼脂糖凝胶电泳的步骤以及其它重组DNA的标准技术在Sambrook等，分子克隆实验手册，第二版，Cold Spring HarborLaboratory Press，Cold Spring Harbor，NY(1989)中作了描述。按照生产商提供的步骤，质粒DNA被核酸内切酶消化。酶是从New England Biolabs(Beverly，MA)，Life Technologies(Rockville，MD)或Promega(Madison，WI)获得。限制性消化在40mM tris-醋酸盐，1mM EDTA缓冲液中，通过0.8％ W/V琼脂糖电泳进行分析。片段的大小是通过与已知分子量(1Kb梯度，Life Technologies，Rockville，MD)的DNA片段作比较后确定的。
C.杂交探针的制备杂交探针在限制性消化后从质粒中被分离出来，或是通过所述聚合酶链反应产生。依照生产商提供的步骤，探针被放射性物质标记为具有特异的放射性，所述放射性标记采用的是New England Nuclear(Boston，MA)的EasyTidesTMα32P-dCTP(3000Ci/mmol)以及Amersham Pharmacia Biotech(Piscataway，NJ)的rediprimeTMII随机引物标记系统。
杂交探针被用以识别含有Fα生物合成基因簇的粘粒(来自蓝灰链霉菌菌株142以及野生型蓝灰链霉菌粘粒文库)，确定和识别转化结合子和切除体(excisants)，并且有助于产生Fα生物合成基因簇的精确的限制性图谱，从而确定基因身份。不是PCR扩增反应所制备的，就是从所述克隆中剪切获得的这些杂交探针，总结于下表1中。
表1
质粒的分离、维持以及繁殖A.质粒的分离未经转化的以及用此处所述的载体转化的大肠杆菌菌株，均采用与Sambrook等，分子克隆实验手册，第二版，Cold Spring Harbor Laboratory Press，Cold Spring Harbor，NY(1989)中所描述的类似的十分确定的方法培养。
质粒DNA通过使用从QIAGEN(Valencia，CA)获得的试剂和材料从大肠杆菌培养物中被分离出来。根据待分析的菌株的数量，所使用的miniprep质粒分离系统包括QIAprep_Spin Miniprep试剂盒(用以从数量相对少的菌株中的质粒分离)；QIAprep_8 Turbo Miniprep试剂盒(用以从数量相对大的菌株中的更高生产量的质粒分离)；或是QIAprep_96 Turbo Miniprep试剂盒(用以从96孔板上的菌株中的部分自动化分离)。为了更大量的从大肠杆菌中分离质粒DNA，使用了QIAGEN Plasmid Midi(最大为100μg)和Maxi(最大为500μg)试剂盒中的试剂和原料或是Clontech(Palo Alto，CA)的Nucleobond AX-100(等于100μg)试剂盒中的试剂和原料。
B.采用质粒DNA转化大肠杆菌质粒DNA经电穿孔转化到大肠杆菌菌株的电感受态细胞或是通过与Sambrook等，分子克隆实验手册，第二版，Cold Spring Harbor Laboratory Press，Cold Spring Harbor，NY(1989)所描述的类似的十分确定的方法，热激而转化到经化学处理的大肠杆菌感受态细胞菌株中。采用适当的抗生素筛选转化体，在使用所述方法分离质粒之后，经限制性核酸内切酶消化之，质粒通过再次使用Sambrook等，分子克隆实验手册，第二版，Cold Spring Harbor Laboratory Press，Cold Spring Harbor，NY(1989)所描述的十分确定的方法，来表征。
C.由大肠杆菌到蓝灰链霉菌的质粒DNA的接合转移在所有的例子中，目的质粒首先经由所述电穿孔方法被转化到命名为ETS12567(pUZ8002)的大肠杆菌菌株中。该菌株是cmr，tetr，dam-，和dcm-。此外，作为质粒pRK2(参见R.Meyer等，Science 1901226-1228(1975))的oriT-型的pUZ8002获得了kanr。经转化的细胞在适当的抗生素选择剂的存在下被培养，所述选择剂包括5μg/ml卡那霉素和100μg/ml阿泊拉霉素。由大肠杆菌转化子到蓝灰链霉菌的质粒DNA的接合转移是通过以下步骤完成的，二者都是在M.Bierman等，Gene11643-49(1992)所描述的步骤的基础上改良的。
接合方法#1将单个经完全分离的转化的大肠杆菌菌落接种于加入了5μg/ml卡那霉素，5μg/ml氯霉素，50μg/ml阿泊拉霉素的3ml LB培养基中，培养物在37℃下以220rpm振荡培养16小时。将100μl冷冻保存的蓝灰链霉菌菌丝体片段接种于10ml TSB(27.5g/L胰蛋白酶大豆肉汤，5g/L酵母提取物，5g/LKH2PO4，pH7.0，经高压灭菌后，加入了100ml/L的20％(w/v)无菌葡萄糖溶液)培养基中，培养物在31℃下以220rpm振荡培养16小时。次日，将100μl的大肠杆菌过夜培养物加入添加了50μg/ml阿泊拉霉素的10ml LB培养基。同时，将2ml的蓝灰链霉菌过夜培养物在含有无菌玻璃珠的管中震荡两分钟。其悬浮液经超声波处理(3X，100％功率下5秒脉冲)；1ml菌丝体片段悬浮液被转移到9mlTSB(27.5g/L胰蛋白酶大豆肉汤，5g/L酵母提取液，5g/L KH2PO4，pH7.0，经高压灭菌后，加入了100ml/L的20％(w/v)无菌葡萄糖溶液)中。两种培养物在37℃下以220rpm振荡培养，直到大肠杆菌培养物的600nm的光吸收值达到0.4-0.6为止。每种培养物中的细胞通过离心收集，用LB洗两遍，然后悬浮于500μl 2XYT(16g/L胰蛋白胨，10g/L酵母提取物，5g/L NaCL，pH7.0)。将两种制备物等量(100μl)混合；混合物在50℃下培养5分钟；然后通过离心收集细胞。移去上清液后，将细胞沉淀悬浮于100ml的2XYT(16g/L胰蛋白胨，10g/L酵母提取物，5g/L NaCL，pH7.0)，然后涂布于SFM(25g/L大豆粉营养大豆，25g/L甘露醇，20g/L 琼脂，0.462g/L L-半胱氨酸，0.462g/L L-精氨酸，0.462g/L L-脯氨酸)平板上。这些平板在37℃下培养16小时，然后涂布含有0.5mg萘啶酮酸以及1mg阿泊拉霉素(最终浓度分别为20μg/ml和40μg/ml)的1ml无菌溶液。这些平板在37℃下培养直到菌落完全出现。
接合方法#2将单个完全分离的转化的大肠杆菌菌落接种于加入了5μg/ml卡那霉素，5μg/ml氯霉素，100μg/ml阿泊拉霉素的3ml LB培养基中，培养物在37℃下以220rpm振荡培养16小时。将1ml冷冻保存的蓝灰链霉菌菌丝体片段接种于25ml KB3培养基(10g/L细菌用胰蛋白胨，5g/L酵母提取物，3g/L牛肉提取物，1g/L KH2PO4，1g/L K2HPO4，1.5g/L Difco琼脂，pH6.8，浓度为0.5ml/L含有30g/L FeSO4、30g/L ZnSO4.7H2O、4g/L MnSO4、4g/L CuCl2.5H2O和0.4g/L CoCl2.6H2O的痕量金属溶液)中，培养物在31℃下以220rpm振荡培养16小时。次日，1ml的大肠杆菌过夜培养物接种于加入了50μg/ml阿泊拉霉素的9mlLB培养基中。同时，将5ml的蓝灰链霉菌过夜培养物在含有无菌玻璃珠的管中震荡两分钟。将2.5ml的均一培养物接种至25ml的KB3培养基(10g/L细菌用胰蛋白胨，5g/L酵母提取物，5g/L牛肉提取物，1g/L KH2PO4，1g/L K2HPO4，1.5g/L Difco琼脂，pH6.8，浓度为0.5ml/L含有30g/L FeSO4、30g/LZnSO4.7H2O、4g/L MnSO4、4g/L CuCl2.5H2O和0.4g/L CoCl2.6H2O的痕量金属溶液)中，两种培养物在37℃下培养3小时。每种培养物中的细胞通过离心收集，并用水洗两遍。然后，将大肠杆菌和蓝灰链霉菌的细胞沉淀分别悬浮于1ml和2ml的TSB(27.5g/L胰蛋白酶大豆肉汤，5g/L酵母提取物，5g/L KH2PO4，pH7.0，经高压灭菌后，加入了100ml/L的20％(w/v)无菌葡萄糖溶液)。10μl蓝灰链霉菌悬浮液和100μl大肠杆菌悬浮液与890μl TSB(27.5g/L胰蛋白酶大豆肉汤，5g/L酵母提取物，5g/L KH2PO4，pH7.0，经高压灭菌后，加入了100ml/L的20％(w/v)无菌葡萄糖溶液)混合，然后将100μl混合液涂布于加入了10mM MgCl2的AS-1(1g/L酵母提取物，0.2g/L L-丙氨酸，0.2g/L L-精氨酸，0.5g/L L-天冬酰胺，5g/L可溶性淀粉，2.5g/L NaCL，10g/L Na2SO4，20g/L琼脂，pH7.5)平板上。这些平板在37℃下培养16小时，然后涂布3ml R2琼脂(100g/L蔗糖，10g/L葡萄糖，10g/L MgCl2，0.25g/L H2SO4，0.1g/L酪蛋白氨基酸，25g/L琼脂)。在使用中，以下溶液被加入到每个装有R2琼脂的80ml烧瓶中1ml 0.5％ K2HPO4；8ml 3.68％ CaCl2.2H2O；1.5ml 20％ L-脯氨酸；10ml 5.73％ TES，pH7.2；0.5ml 1NNaOH；以及d 1ml含有40mg/L ZnCl2，200mg/L FeCl3.6H2O，10mg/L CuCl2.2H2O，10mg/L MnCl2.4H2O，10mg/L Na2B4O7.10H2O，10mg/L(NH4)6Mo7O24.4H2O)的痕量元素溶液。该溶液还加入了100μg/ml的阿泊拉霉素和100μg/ml的萘啶酮酸(最终浓度)。这些平板在37℃下培养直到菌落完全出现。
采用上述任一方法，推定的转化结合子被反复挑选接种于含有100μg/ml的阿泊拉霉素和100μg/ml的萘啶酮酸的新鲜平板上，直到由作为质粒来源的大肠杆菌菌株导致的可见的污染被治愈。
编码用于生产LL-F28249化合物的完整生物合成途径的，来自蓝灰链霉菌非产蓝亚种的经纯化的DNA，与本专利申请有关根据37 C.F.R.§1.808规定的条件，已经依据布达佩斯条约被保藏于美国典型培养物保藏中心(ATCC)，10801University Boulevard，Manassas，Virginia 20110-2209，U.S.A。更明确的是，经纯化的粘粒DNA，在此完全和明确的描述为Cos11，Cos36 and Cos40，已于2002年5月24日保藏于ATCC，指定的ATCC专利保藏号分别为PTA-4392，PTA-4393和PTA-4394。应该重视的是，相关的经纯化的DNA，和那些容易采用定点突变和此处所述技术构建的其他的粘粒，或其它含有相关核苷酸序列的质粒或粘粒也包括在本发明的范围内。
下面的实施例显示了本发明的某些方面。然而，应该认识到这些例子只是为了阐明，而并不是旨在对于本发明的条件和范围的完全限定。应该重视的是，当给定典型的反应条件(例如温度，反应时间等等)，尽管一般说来稍有不便，但是反应条件的特定范围的上下区域也是可以采用的。实施例都是在室温(大约23℃至大约28℃)条件下以及大气压下进行的。除非特别提到，这里所指的部分和百分数都是基于重量基础，并且所有的温度都是用摄氏温度来表示的。
对于本发明的进一步理解可以通过下面对本发明不具限制性的实施例来获得。
实施例1制备LL-F28249化合物的生物合成基因簇的表征A.含有Fα生物合成基因簇的粘粒的分离和表征1.蓝灰链霉菌粘粒文库的构建基因组DNA采用在D.A.Hopwood等在链霉菌的遗传操作实验手册，JohnInnes Foundation Press，Norvich，UK(1985)(“分离链霉菌的“总”DNA步骤3)中所述方法，从蓝灰链霉菌(野生型以及称作142的Fα生产菌株)中分离出来。接着制备出的蓝灰链霉菌基因组DNA如下通过限制性核酸内切酶Sau3AI进行部分消化。制备含有Sau3AI以及基因组IDNA的混合物，在一定的时间点((0，5，10，15，20，30，以及45分钟)，各取出一部分，并且通过加入EDTA至终浓度为10mM来终止反应。从每个时间点终止反应的反应物中取出一部分在0.3％w/v琼脂糖中在25伏电压下电泳16小时分离。选择那些含有大小主要介于23Kbp至50Kbp的DNA片段的反应时间点从而构建粘粒文库。同时，pSuperCos 1(Stratagene，La Jolla，CA)采用限制性核酸内切酶XbaI消化；用小牛肠碱性磷酸酶脱去磷酸；然后在经过乙醇沉淀之后，该线性质粒通过限制性核酸内切酶BamHI消化，从而除去一个COS位点。依照生产商提供的步骤，蓝灰链霉菌基因组DNA的Sau3AI片段被连接到线性的经BamHI消化的pSuperCos 1中。用Gigapack_III XL Packaging Extract将所得到的重组粘粒DNA制备产物包装，在用氯仿溶解所得到的λ噬菌体颗粒后，粘粒文库被转化至大肠杆菌VCS257。这些操作都是采用生产商(Stratagene，La Jolla，CA)提供的试剂、原料以及步骤进行的。
2.含有Fα生物合成基因簇的粘粒的分离基因组DNA采用在D.A.Hopwood等在链霉菌的遗传操作实验手册，JohnInnes Foundation Press，Norvich，UK(1985)(“分离链霉菌的“总”DNA步骤3)中所述方法从除虫链霉菌中分离得到的。该基因组DNA的制备产物，作为模板，用于通过聚合酶链反应扩增除虫菌素生物合成基因簇的组件1酮脂酰合成酶酶域的区域。所使用的寡核苷酸探针是根据已保藏在公共数据库中的除虫菌素生物合成基因簇的核苷酸设计的。通过与除虫菌素酮脂酰合成酶探针杂交，筛选蓝灰链霉菌142菌株的粘粒文库的菌落，从而分离得到超过30个潜在含有I型聚酮合成酶DNA的粘粒。最初，这些粘粒在经BamHI消化之后，通过琼脂糖凝胶电泳、用除虫菌素组件1酮脂酰合成酶探针进行Southem杂交以及限制性核苷酸序列分析来进行分析。将这些数据与由MacNeil及其同事(参见D.J.MacNeil等，Gene 115119-125(1992)和D.J.MacNeil等，Annals of the NewYork Academy of Sciences 721123-132(1994))报道的数据相比较，显示这些粘粒中的两个(称作Cos7和Cos11)跨越了Fα生物合成基因簇的主要部分。由MacNeil及其同事所提供的有限的数据也被作为原始的依据以支持包含大部分细件3的5.7Kbp的NotI-EcoRI片段的分离。一个这种5.7Kbp的NotI-EcoRI片段的克隆被制备出来(称为pNE57)。这个5.7Kbp的片段的核苷酸序列被全部测定。然后该Fα生物合成基因簇(来自从Fα生产菌株中分离的基因组DNA)的片段被用作探针来筛选野生型蓝灰链霉菌粘粒文库，从而分离得到45个潜在含有I型聚酮合成酶DNA的粘粒。通过使用所述方法，绘制出这些粘粒的BamHI，NotI以及EcoRI限制性酶切图谱。在比较这些限制性图谱与由MacNeil及其同事报道的不完全的数据的基础上，识别出两个呈现跨越了Fα生物合成基因簇的主要部分的粘粒(来自野生型菌株，称为Cos36和Cos40)。
为了鉴别跨越了Fα生物合成基因簇的“末端”，但并不含有核心聚酮合成酶DNA的重要的区域的粘粒，采用了下述策略。分离自Cos11(源自蓝灰链霉菌菌株142)的5.5Kbp的BamHI片段被用作探针，对已经筛选过的野生型蓝灰链霉菌粘粒再进行筛选，以识别出可将基因簇向“左”延伸的粘粒。许多与该探针杂交的粘粒被识别，在制作限制性图谱之后，这些粘粒中的一个，COS14，被确定支持基因簇延伸至左边最远。分离自Cos36的3′端的，一个500bpNotI片段，被用作探针对野生型蓝灰链霉菌粘粒文库再进行筛选，以识别出可将基因簇向“右”延伸的粘粒。许多与该探针杂交的另外的粘粒被确定，在制作限制性图谱之后，这些粘粒中的一个，COS50，被确定支持基因簇延伸至右边最远。
3.含有Fα生物合成基因簇的粘粒的限制性图谱起先，来自蓝灰链霉菌菌株142粘粒文库的，与除虫菌素酮酯酰合成酶探针杂交的超过30个的粘粒，以及来自野生型蓝灰链霉菌粘粒文库的，与Fα组件3探针(pNE57)杂交的超过30个的粘粒，在经由BamHI，NotI和EcoRI消化后，绘制出限制性酶切图谱。在这种初步分析的基础上，以及在将该限制性图谱与由MacNeil以及其同事(参见D.J.MacNeil等，Gene 115119-125(1992)and D.J.MacNeil等，Annals of the New York Academy of Sciences 721123-132(1994)提供的不完全的数据比较的基础上，几个粘粒被筛选出来以做进一步的分析。这些粘粒(来自蓝灰链霉菌菌株142的Cos7和Cos11；来自野生型蓝灰链霉菌的Cosl2，Cosl4，Cos36，Cos40和Cos50)在经过由BamHI，NotI和EcoRI消化，以及BamHI/MYuI，NotI/FcoRI，BamHI/EcoRI，SacI/EcoRI，和NotI/MluI的双酶切后，其限制性酶切图谱被仔细地绘制出来。为了解决所观察到的限制性图谱中的含糊之处，这些粘粒的亚克隆如下表2所总结的被构建出来，这些亚克隆已经如同前面所述被详尽地绘制了酶切图谱。
表2
<p>表5
基于广泛的限制性图谱显示的跨越Fα生物合成基因簇的主要部分的两个粘粒从野生型蓝灰链霉菌粘粒文库中被分离出来。这些粘粒通过末端测序如前面所述制备的随机的，按大小筛选的粘粒DNA子文库而被完整测序。此外，由许多亚克隆的不同插入子(如下表3)所制备的随机、大小筛选的子文库也被测序。最后，产生出来的，以支持Fα生物合成基因簇完全限制性酶切图谱的亚克隆的主要部分也通过通用引物进行了末端测序。
表3
2.用以核苷酸序列分析的子文库的构建为了在粘粒以及衍生于这些粘粒的亚克隆中产生大量的插入片段，需要大量的质粒DNA。在培养基(一般是1L)中接种目的克隆，在37℃下培养过夜。通过使用包括在QIAGEN Plasmid Midi(总计100μg)以及Maxi(总计500μg)试剂盒，或包括在Clontech(Palo Alto，CA)的Nucleobond AX-100(总计100μg)试剂盒中的试剂和原料，质粒(粘粒)DNA从这些培养物中被分离出来。这些质粒(粘粒)中的插入片段通过适当的限制性核酸内切酶消化被剪切，所得片段经0.8％w/v的琼脂糖电泳分离。目的片段从这些凝胶中剪切得到，并采用包含在QIAEX II_(适于片段大于10Kbp)或QIAquick II(适于片段小于10Khp)GelExtraction Systems from QIAGEN(Valencia，CA)中的试剂原料以及步骤分离出这些条带中的DNA。然后，通过使用超声波细胞破碎器以10％的功率，用超声波对DNA进行随机剪切。优化超声处理的时间，从而产生目的大小的片段(一般对于从粘粒中分离的大的插入片段需大约18秒，对于从粘粒的质粒亚克隆中分离的小的插入片段需大约8秒)。经乙醇沉淀之后，在含有2.5μl的10×T4 DNA聚合酶链反应缓冲液，1μl的25μg/ml BSA，以及1.5μl的T4 DNA聚合酶的25μl反应体积中，采用T4 DNA聚合酶(New England Biolabs，Beverly，MA)“钝化”DNA片段。反应混合物在16℃下温育20分钟，然后通过0.8％w/v的琼脂糖电泳分离。含有大小在1.5Kbp至2.5KbpDNA的凝胶区域被切下，采用包含在QIAquick II Gel Extraction System from QIAGEN(Valencia，CA)中的试剂原料和步骤，从凝胶中提取出DNA。经纯化的DNA通过乙醇沉淀，并重悬于8μl水中。接着，通过使用生产商(Invitrogen，Carlsbad，CA)提供的试剂原料和步骤，这些DNA片段被克隆到pCR_-Blunt上，所得的连接产物被转化到经化学处理的感受态大肠杆菌TOP 10中。挑选克隆，并将其接种于96孔深孔培养板中，其中加入了含50μg/ml卡那霉素的2ml LB培养基。采用QIAprep_96 TurboMiniprep Kits试剂盒中的试剂原料和步骤，纯化得到每一培养物中的质粒DNA。尽管有插入片段的克隆的发生频率超过90％，但仍采用EcoR I消化每一个质粒，且所得片段通过0.8％w/v的琼脂糖电泳分离，以证实是否存在所需大小的插入片段。那些确实含有所需插入片段的克隆通过所述的通用引物被测序。
3.生物合成的组件以组件中的酶域的鉴定许多组件聚酮化合物生物合成基因簇已经被表征和操作。此外，大量组件聚酮化合物生物合成基因簇的核苷酸序列已经保存在公众数据库中。一般说来，组件聚酮化合物生物合成基因簇中的组件以及这些组件中的酶域能够通过对公众数据库进行BLAST检索所确定，这些公众数据库的广泛的使用有助于当前对Fα生物合成基因簇(参见S.F.Altschul等，Nucleic Acids Research253389-3402(1997))的分析。此外，采用了一篇最近总结的有关识别组件聚酮合成酶酶域的方法的参考文献(S.J.Kakavas等，Journal of Bacteriology1797515-7522(1997))，特别是该文献描述了丙二酰类酰基转移酶酶域与甲基丙二酰类酰基转移酶酶域之间的差别。Leadlay和其同事最早描述了区别丙二酰类酰基转移酶酶域与甲基丙二酰类酰基转移酶酶域的方法(参见T.Schwecke等，Proceedingsofthe National Academy of Sciences USA927839-7843 (1995))。
下面的表4阐明了对于五个开放阅读框的描述，这五个开放阅读框一起编码聚酮合成酶的载入域以及13个组件。对于每一个开放阅读框，其在Fα生物合成基因簇的位置(核苷酸上)以及预期的基因产物的长度(氨基酸)被阐明。此外，在预期的基因产物(在氨基酸)内的每一个生物合成域的大致位置也被阐明。所用缩略语如下表示ACP，酰基载体蛋白；ATm，丙二酰类酰基转移酶；Atmm，甲基丙二酰类酰基转移酶；DH，脱水酶；ER，烯酰还原酶；KR，酮还原酶KS，酮脂酰合成酶；LD，载入域；TE，硫酯酶。
表4ORF4nt 12850-19875(2339 aa)ORF9nt 52809-69833(5675 aa)名称载入域-Mod1 名称Mod8-Mod10ATmm-LD aa 22-350KS-8 aa 39-465ACP-LD aa 365-450 ATmm aa 574-904KS-1aa 473-897 DH-8 aa 926-1106ATmm-1 aa 1006-1339 ER-8 aa 1366-1718DH-1aa 1359-1547 KR-8 aa 1726-1908KR-1aa 1865-2052 ACP-8 aa 1995-2080ACP-1 aa 2137-2223 KS-9 aa 2102-2529ATm-9 aa 2661-2986ORF5nt 19865-31036(3724 aa)DH-9 aa 3009-3188名称Mod2-Mod3 KR-9 aa 3492-3674ACP-9 aa 3753-3842KS-2aa 34-466KS-10 aa 3864-4290ATmm-2 aa 574-908 ATmm-10 aa 4402-4732KR-2aa 1211-1391 DH-10 aa 4753-4928ACP-2 aa 1473-1559 KR-10 aa 5234-5416KS-3aa 1578-2005 ACP-10aa 5499-5586ATm-3 aa 2136-2476DH-3aa 2486-2667 ORF10nt69929-85429(5167aa)ER-3aa 2925-3279 名称Mod11-Mod 13KR-3aa 3287-3466ACP-3 aa 3556-3640 KS-11 aa 34-456ATm-11aa 578-916ORF6nt 31115-49246(6044 aa)KR-11 aa 1199-1380名称Mod4-Mod7 ACP-11aa 1464-1549KS-12 aa 1570-1996KS-4aa 34-456ATmm-12 aa 2105-2442ATm-4 aa 582-907 KR-12 aa 2724-2906ACP-4 aa 950-1031 ACP-12aa 2992-3076KS-5aa 1055-1481 KS-13 aa 3096-3519ATm-5 aa 1613-1938 ATm-13aa 3631-3975KR-5aa 2247-2427 DH-13 aa 4003-4188ACP-5 aa 2516-2601 KR-13 aa 4505-4687KS-6aa 2621-3047 ACP-13aa 4780-4866ATm-6 aa 3168-3493 TE-13 aa 4893-5167KR-6aa 3802-3983ACP-6 aa 4078-4164KS-7aa 4189-4615ATmm-7 aa 4727-5056DH-7aa 5078-5257KR-7aa 5588-5768ACP-7 aa 5868-5952
4.其它生物合成途径基因的鉴定不论被发现的密集在核心聚酮合成酶基因周围的其它开放阅读框是否在Fα生物合成中起作用，即使有作用，那种作用是什么也是基于那些开放性阅读框的核苷酸以及预测的氨基酸序列与已保存在公众数据库中的序列的BLAST比较得出的(参见S.F.Altschul等，Nucleic Acids Research 253389-3402(1997))。通过这些方法，初步鉴定出至少6个其它的基因涉及Fα生物合成。
在下面的表5中对这6个额外的，可能编码涉及Fα生物合成的基因的开放阅读框做了描述。对于每一个开放阅读框，其在Fα生物合成基因簇的位置(核苷酸上)以及预测的基因产物的长度(氨基酸)被阐明。此外，这里还包括了每一个开放阅读框的用于明确其在Fα生物合成中的推定作用的BLAST结果的简介。
表5ORFAnt 382-2514(711 aa)名称K+-转移腺苷三磷酸酶亚基B(与Fα生物合成基因簇无关)ORFBnt 2511-4175(555 aa)名称K+-转移腺苷三磷酸酶亚基A(与Fα生物合成基因簇无关)ORF1nt 7697-10465(923 aa)名称调节蛋白ORF2nt 10791-11570(260 aa)名称硫酯酶ORF3nt 11659-12462(268 aa)名称还原酶ORF7nt 50449-51303(285 aa)名称甲基转移酶ORF8nt 51300-52706(425 aa)名称p450ORF11nt 85574-86338(254 aa)名称氧化还原酶
ORFXnt 87037-88293(419aa)名称内-1，3-β-葡萄糖苷酶(与Fα生物合成基因簇无关)ORFA和ORFBBLAST结果显示，在ORFA和ORFB之间，以及K+-改变位置三磷酸酰苷酶亚基B和A之间，分别存在相当大的同源性，尤其是结合杆菌基因(其核苷酸序列被直接递交给公众数据库)。这些基因都与Fα生物合成基因簇无关。
ORF 1BLAST结果显示，在核苷酸水平，ORF 1是与来自委内瑞拉链霉菌(S.venezuelae)(参见Y.Xue等，Proceedings of the National Academy of SciencesUSA 9512111-12116(1998))的大环内酯生物合成基因簇pikCD操纵子的推定转录激活子以及来自雷帕霉素产生生物吸水链霉菌(S.hygroscopicus)(参见X.Ruan等，Gene 2031-9(1997))的I型聚酮合成酶生物合成基因簇的一个推定的调节蛋白相关。在预期的氨基酸水平，基因产物呈现出与大肠杆菌narL基因产物相关的推定的转录激活子家族的有限的同源性。基于BLAST结果，ORF 1呈现出编码一个转录激活子。
ORF2BLAST结果显示，ORF2与硫酯酶在核苷酸以及氨基酸序列水平的显著同源性，所述硫酯酶包括地中海拟无枝酸菌(Amycolatopsis mediierranei)利福霉素生物合成基因簇(参见P.R.August等，Chemistry & Biology 569-79(1998))中的硫酯酶，以及灰色链霉菌S.griseus杀假丝菌素生物合成基因簇(参见L.M.Criado等，Gene 126135-139(1993))中的硫酯酶。基于这些BLAST结果，ORF2呈现出编码一种硫酯酶。
ORF3对BLAST结果分析显示，ORF3与产蓝链霉菌(S.cyanogenus)S136竹桃霉素生物合成基因簇(参见L.Westrich等，FEMS Microbiological Letters170381-387(1999))中的还原酶同源。在预期的氨基酸序列水平，BLAST结果显示出ORF3基因产物与负责Aspergillus parasiticus黄曲霉毒素生物合成途径(参见C.D.Skory等，Applied and Environmental Microbiology 583527-3537(1992))中的杂色曲菌素A至柄曲霉素转化的氧化还原酶之间的同源性。基于这些BLAST的结果，ORF3呈现出编码一种还原酶。
ORF7BLAST结果显示，ORF7与甲基转移酶在核苷酸水平的显著同源性，所述甲基转移酶包括在淡紫灰链霉菌(S.lavendulae)丝裂霉素C生物合成基因簇(参见Y.Q.Mao等，Chemistry & Biology 6251-263(1999))中的甲基转移酶，以及红色糖多孢菌(Saccharopolyspora erythraea)红霉素生物合成基因簇(参见S.F.Haydock等，Molecular和General Genetics 230120-128(1991))中的甲基转移酶。基于这些BLAST的结果，ORF7呈现出编码一种甲基转移酶。
ORF8BLAST结果显示，ORF8与推定的细胞色素P450′s之间的有限的同源性。所述细胞色素P450′s包括在玫瑰暗黄链霉菌(S.Roseofulvus)富伦菌素生物合成基因簇(参见C.D.Reeves和C.L.Soliday，direct submission)的P450′s，以及始旋链霉菌(S.Pristinaespiralis)原始霉素生物合成基因簇(参见V.deCrecy-Lagard等，Journal of Bacteriology179705-713(1997))中的P450′s。在预期的氨基酸序列水平，ORF8显示出与哺乳动物细胞色素P450′s的一大家族的同源性。基于这些BLAST的结果，ORF8呈现出编码细胞色素P450。
ORF11BLAST结果显示，ORF11与氧化还原酶在核苷酸以及氨酸序列水平的显著同源性，所述氧化还原酶包括在紫灰链霉菌(S.violaceoruber)粒菌素生物合成基因簇(D.H.Sherman等，EMBO Joumal 82717-2725，(1989))中的氧化还原酶，以及肉桂链霉菌(S.Cinnamonensis)莫能菌素生物合成基因簇(参见V.de Crecy-Lagard等，Journal of Bacteriology179705-713(1997))中的氧化还原酶。基于这些BLAST的结果，ORF11呈现出编码一种氧化还原酶。
ORF XBLAST结果显示ORFX与来自溶黄嘌呤厄氏菌(Oerskoviaxanthineolvtica)(参见S.H.Shen等，Journal of Biological Chemistry2661058-1063(1991))的葡聚糖内-1，3-β-葡糖苷酶的同源性。该基因与Fα生物合成基因无关。
在经表征的ORFB和ORF1之间的3.5Kbp区域有一些开放阅读框，其基于核苷酸序列特征(G+C含量，潜在的核糖体结合位点)编码蛋白质。然而，BLAST分析并没有显示出推定的蛋白质的预期的氨基酸序列与已经保存在公众数据库中的蛋白质的序列之间存在显著的同源性。因此，不可能仅仅基于它们的核苷酸(或预期的氨基酸)序列，从而确定那些Fα生物合成中的推定蛋白质的功能。此外，在经表征的ORFX和现在已经获得的核苷酸序列的末端之间的7.8Kbp区域，存在许多开放阅读框。由于ORFX编码的是在Fα生物合成中看似没有作用的的基因，并且由于大环内酯生物合成基因通常是丛生的，由超出ORFX的开放性读框编码的假定的蛋白质并不参与Fα生物合成。
实施例2基因置换，整合体(Integrants)以及切除体(excisants)的表征A.基因置换为了培养一种能够直接发酵产生23-酮-Fα的蓝灰链霉菌菌株，产生Fα产物的衍生物，寻求其中的组件3酮还原酶被没有功能的变异体置换的菌株。采用如下方法设计出一系列的定向氨基酸取代，每一设计都阻断酮还原酶的活性并最低限度影响余下的聚酮合成酶。当将蓝灰链霉菌Fα生物合成基因簇的组件3酮还原酶酶域的预期的氨基酸序列与大量生物活性酮还原酶酶域的预期的氨基酸序列比对时，产生一多重氨基酸序列比对。这些酮还原酶酶域序列来自除虫链霉菌(S.Avermitilis)除虫菌素生物合成基因簇，红色糖多孢菌红霉素生物合成基因簇，吸水链霉菌雷帕霉素生物合成基因簇，天青链霉菌S.caelestis尼达霉素生物合成基因，以及地中海拟无枝酸菌利福霉素生物合成基因簇。已知的无功能的三个酮还原酶酶域(来自红色糖多孢菌的红霉素生物合成基因簇的组件3、天青链霉菌尼达霉素生物合成基因簇的组件4以及地中海拟无枝酸菌的利福霉素生物合成基因簇的组件3的所谓的“隐藏的”酮还原酶酶域)也包括在该序列比对中。这种多重氨基酸序列比对，易于支持对常见于大部分的生物活性酮还原酶酶域但是在无功能还原酶酶域缺乏(或改变)的相对无变异氨基酸序列的鉴定。
如下所述，通过同源重组，可以对蓝灰链霉菌进行基因置换，从而使Fα生物合成基因簇的组件3酮还原酶酶域的目的变异体可以被所述组件3酮还原酶酶域的工程变异体所置换。
1.定点位点诱变的质粒的构建QuikchangeTM定点位点诱变步骤是一个基于聚合酶链反应的双链方法，该方法需要两条诱变的寡核苷酸，分别对应于相应于DNA双链区域的每一条。当使用大质粒，尤其是含有高G+C含量DNA的大质粒时，该方法有效性降低。因此，Fα的组件3酮还原酶酶域的定点位点诱变在命名为pKR0.9(参见图3)的载体中进行，发生在pSL301(Invitrogen，Carlsbad，CA)的BstEII-AatII位点，为pNE57的900bp的BstEII-AatII片段(并含有目的区域组件3酮还原酶酶域)。
2.定点位点诱变采用QuikChangeTM的定点位点诱变试剂盒(Stratagene，La Jolla，CA)的生产商提供的试剂原料和步骤，进行定点位点诱变，产生了5个Fα组件3酮还原酶酶域的变异体。接下来的氨基酸取代通过使用下面的突变的寡核苷酸在pKR0.9中产生。
“179”GGTGTLG(SEQ ID NO13)至GAASTLG(SEQ ID NO14)5’-CTGGTGACGGGCGCTGCAAGCACTCTGGGGGCG(SEQ ID NO15)3’-GACCACTGCCCGCGACGTTCGTGAGACCCCCGC(SEQ ID NO16)“204”LVSRRGM(SEQ ID NO17)至LVAAAGM(SEQ ID NO18)5’-GCGGCATCTGCTGCTGGTGGCAGCGGCAGGCATGGCCGCCGCCGGTG(SEQ IDNO19)3’-CGCCGTAGACGACGACCACCGTCGCCGTCCGTACCGGCGGCGGCCAC(SEQ IDNO20)“260”HTAGVLD(SEQ ID NO21)至HTPPLLD(sEQ ID NO22)5’-GACCGCTGTGGTGCACACGCCACCTCTCCTGGACGACGCCACCGTG(SEQ IDNO23)3’-CTGGCGACACCACGTGTGCGGTGGAGAGGACCTGCTGCGGTGGCAC(SEQ IDNO24)“283”GAKVD(SEQ ID NO25)至GAAVD(SEQ ID NO26)5’-GATGCGGTGCTCGGGGCGGCTGTGGACGGTGCCCTGCAC(SEQ ID NO27)3’-CTACGCCACGAGCCCCGCCGACACCTGCCACGGGACGTG(SEQ ID NO28)“306”VLFSSAA(SEQ ID NO29)至VLFAAAA(SEQ ID NO30)5’-GTCGGCGTTCGTGCTGTTCGCAGCGGCCGCCGGGGTCCTGG(SEQ ID NO31)3’-CAGCCGCAAGCACGACAAGCGTCGCCGGCGGCCCCAGGACC(SEQ ID NO32)QuikChangeTM变反应在50μl的最终反应体积中进行，其中含有125ng的每一突变核苷酸，50ng的pKR0.9，0.7μl的Pfu DNA聚合酶以及2.5％DMSO。反应进行22个循环的扩增(95℃ 45秒，63℃ 1分钟，以及70℃ 10分钟)，按照生产商提供的详尽的步骤克隆扩增的产物。在完成了定点诱变步骤之后，挑取克隆，并接种于加入了100μg/ml羧苄青霉素的2ml LB培养基中。通过使用QIAprepOR 8 Turbo Miniprep Kits中的试剂原料和步骤，从每一个培养物中纯化质粒DNA。Fα的组件3酮还原酶酶域的突变的900bp的BstEII-AatII区域被完全测序以确认所需的变化确已发生。
3.用于整合的质粒的构建一种三步连接被用来将Fα的组件3酮还原酶酶域的5个定点位点突变体与侧翼序列组合，以促进采用pKC1132为骨架的同源整合。这三种成分包括4.3Kbp的pNE57(含有邻近突变区的Fα的组件3的主要部分)的Not I-BstE II片段；1.1Kbp的6个pKR0.9构建体(含有Fα的组件3酮还原酶酶域的5个定点位点突变以及野生型Fα的组件3酮还原酶酶域)的BstEII-PstI片段；3.6Kbp的pKC 1132(含有所有在大肠杆菌和链霉菌中所得质粒的筛选和复制所必须的元件))的PstI-NotI片段。这些操作导致了pFDmod3/5.2质粒系列的产生。这些质粒然后被用以构建用以整合的大约1Kbp侧翼DNA被移除的质粒。这些质粒通过采用EcoRI消化每一pFDmod3/5.2质粒来构建。在pKC 1132中，该EcoRI位点紧邻着NotI位点，所述pKC1132被用以引入4.3Kbp的pNE57(含有Fα的组件3的主要部分)的NotI-BstEII片段。在标准反应条件下，使用T4DNA聚合酶，补平3′突出端。线性化的质粒被MscI消化。所得消化产物通过0.8％w/v琼脂糖电泳分离，将所需片段从凝胶中切出，通过使用QIAGEN(Valencia，CA)的QIAquickII Gel Extraction System中的试剂原料和步骤，提取DNA。纯化的DNA通过乙醇沉淀被收集，连接产生pFDmod3/4.2质粒系列(参见图5)。
pFDmod3/5.2质粒系列(参见图4)以及pFDmod3/4.2质粒系列(参见图5)的质粒通过所述方法被转化到大肠杆菌ETS 12567(pUZ8OO2)中。然后，根据所述的方法，这些经转化的大肠杆菌菌株被用作接合转移蓝灰链霉菌的DNA的来源。
4.蓝灰链霉菌转化结合子以及切除体(excisants)的基因细DNA的分离和分析来源于D.A.Hopwood等在链霉菌的遗传操作实验手册，John InnesFoundation Press，Norvich，UK(1985)(“链霉菌的分离”“整个”DNA；步骤3)中所述方法的一种改良方法被用以从蓝灰链霉菌中分离少量基因组DNA。挑取推定的蓝灰链霉菌的转化结合子以及切除体(excisants)，并且接种于3ml KB3培养基中(10g/L细菌用胰蛋白胨，5g/L酵母提取物，3g/L牛肉提取液，1g/LKH2PO4，1g/L K2HPO4，1.5g/L Difco琼脂，pH6.8以及浓度为0.5ml/L的含有30g/L FeSO4、30g/L ZnSO4.7H2O、4g/L MnSO4、4g/L CuCl2.5H20和0.4g/LCoCl2.6H2O的一种痕量金属溶液)。培养物在31℃下以220rpm振荡培养24-28小时。然后，500μ1等份的该培养物中的细胞通过在微量离心管中13,000rpm转速下离心5分钟，去掉上清液。用水洗涤沉淀，然后将其悬浮于450μl的SET(0.3M蔗糖，25mM EDTA，25mM Tris，pH8.0，含有4mg/ml溶菌酶和50μg/ml RNA酶A)中。悬浮液在37℃下培养2-4小时。加入250μl的2％SDS溶液，样品被震荡1分钟。该样品通过250μl的酚氯仿(1∶1)溶液提取，提取相通过在微量离心管13,000rpm转速下离心5分钟获得分离。水相被转移到一个新的试管中，在加入了1/10体积的3M醋酸钠之后，DNA通过加入等体积的异丙醇得以沉淀。通过在微量离心管13,000rpm转速下1离心5分钟，收集沉淀的DNA，用70％乙醇在-20℃洗涤，然后悬浮于100μl水中。
为了从蓝灰链霉菌中分离更大量的基因组DNA，在25ml KB3培养基(10g/L细菌用胰蛋白胨，5g/L酵母提取物，3g/L牛肉提取液，1g/LKH2PO4，1g/L K2HPO4，1.5g/L Difco琼脂，pH6.8以及浓度为0.5ml/L的含有30g/L FeSO4、30g/L ZnSO4.7H2O、4g/L MnSO4、4g/L CuCl2.5H2O和0.4g/L CoCl2.6H2O的一种痕量金属溶液)中接种目的菌株的菌丝体片段。培养物在31℃下以220rpm振荡培养24-28小时。3ml等份该培养物中的细胞通过在微量离心管13,000rpm转速下离心5分钟，去掉上清液。水洗细胞沉淀后，基因组DNA采用QIAGEN(Valencia，CA)的用以分离总DNA(植物)的DNAeasyTM中的试剂原料和步骤被分离出来。
5.转化结合子的表征推定的转化结合子被涂布于含有100μg/ml阿泊拉霉素，30μg/ml萘啶酮酸，50μg/ml放线菌酮和50μg/ml制霉菌素A的CM琼脂(5g/L玉米浸液，5g/L细菌用蛋白胨，10g/L可溶性淀粉，0.5g/L NaCl，0.5g/L CaCl2·2H2O，20g/L细菌用琼脂)平板上。这些平板在31℃下培养，直到菌落完全出现。然后通过所述方法，从推定的转化结合子中分离出基因组DNA，并根据下述的Southern印迹法以及核苷酸序列分析方法，用于分析。所述等份的基因组DNA制备产物被HindIII/Stu I以及Sal I所消化。所得片段通过0.8％w/v的琼脂糖电泳分离，然后杂交于NytranTM膜上(可通过商业途径由Schleicher & Schuell BioScience，Inc.USA，Keene，NH获得)，并根据与Sambrook等，分子克隆实验手册，第二版，ColdSpring Harbor Laboratory Press，Co1d Spring Harbor，NY(1989)所描述的方法类似的，已确立的方法来进行Southern分析。一般来说，这些Southern印迹与那些通过所述方法制备的mod3-特异探针杂交。所述片段的预期大小是菌株HindIII/StuISalI蓝灰链霉菌生产菌株142 10.8Kbp 4.6Kbp蓝灰链霉菌生产菌株142/pFDmod3/5.2转化结合子 13.3 Kbp4.6Kbp+3.3Kbp蓝灰链霉菌生产菌株142/pFDmod3/4.2转化结合子 12.3 Kbp4.6Kbp+3.3Kbp基于Southern分析的看似正确的转化结合子的目的区域通过标准聚合酶链反应(PCR)被扩增出来。PCR产物被测序以确认获得所需序列。两个引物组合被用来表征这些转化结合子。每一对引物是由一条mod3-特异引物，以及一条载体来源序列的特异引物组成。此外，设计所述引物组，从而一对引物能从“表达盒的右侧”扩增产物，而另一对能够从“表达盒的左侧”扩增产物。所使用的引物对是左 (mod70F)5 ′-TACTGCGCCACACGGAGCCCGAG(SEQ ID NO33)和(P6568B)5′-TGGGTAACGCCAGGGTTTTC(SEQ ID NO34)右 (PECOR1F) 5′-GGAAACAGCTATGACATGATTACG (SEQ IDNO35)和(mod3633B) 5′-TCGGAGCCGCTCCACCTGAG(SEQ ID NO36)以从“正确的”转化结合子分离的基因组DNA作为模板，这些PCR引物能够分别引导6.4Kbp和5.7Kbp产物的扩增。采用下述的寡核苷酸引物，对含有酮还原酶域的这些PCR产物的区域进行测序，以证实所需序列已获得。
″179″转化结合子正向5′-CCTGATGGACGCGGGTGCGC(SEQ ID NO37)反向5′-GACACCGAAACCCCTG(SEQ ID NO38)″204″转化结合子正向5′-CCTGATGGACGCGGGTGCGC(SEQ ID NO39)
反向5′-GCCGTGTGCACCACAGCGGTCA(3(SEQ IDNO40)″260″，″283″，″306″转化结合子正向5′-GTGTGATGTCGCCGACCGCGCCCAGGTC(SEQ IDNO41)反向5′-GCGCTGGTGGGCCAGGGCGTCC(SEQ ID NO42)6.切除体的切除和表征采用所述方法，将通过对PCR产物的Southern分析以及核苷酸序列分析被证实的转化结合子接种于25ml的KB3(10g/L细菌用胰蛋白胨，5g/L酵母提取物，3g/L牛肉提取液，1g/L KH2PO4，1g/L K2HPO4，1.5g/L Difco琼脂，pH6.8以及浓度为0.5ml/L的含有30g/L FeSO4、30g/LZnSO4.7H2O、4g/L MnSO4、4g/L CuCl2.5H2O和0.4g/L CoCl2.6H2O的一种痕量金属溶液)培养基中，培养物在31℃下以220rpm振荡培养48小时。将500μl的培养物转到新鲜的25ml KB3培养基中，在31℃下以220rpm继续进行振荡培养48小时。为了切除事件发生，在没有选择剂的情况下，该过程持续进行许多轮。在3-6轮后，一系列10-1至10-5培养物稀释液被制备出来，250μl的10-3至10-5培养物稀释液等份被涂布到直径为140mm的CM琼脂平板(5g/L玉米浸液，5g/L细菌用蛋白胨，10g/L可溶性淀粉，0.5g/L NaCl，0.5g/L CaCl2·2H2O，20g/L细菌用琼脂)上。这些平板在31℃下培养48-96小时直到菌落完全出现。挑选单个菌落，将其按照双份接种于在CM平板上，CM平板上加入了100mg/ml阿泊拉霉素。这些平板在31℃下培养5天，在这个时间，对于阿泊拉霉素敏感，但能在缺乏选择剂的条件下正常生长的菌落被鉴别出来。然后通过所述方法，从推定的切除体(excisants)中分离出基因组DNA。以基因组DNA制备产物为模板，通过聚合酶链反应(PCR)扩增出目的区域。所用以扩增的引物是(mod70F)5′-TACTGCGCCACACGGAGCCCGAG(SEQ ID NO33)和(mod3633B) 5′-TCGGAGCCGCTCCACCTGAG(SEQ ID NO36)以从“正确的”切除体中分离的基因组DNA作模板，这些PCR引物会引导扩增6.6Kbp的产物。通过下面的寡核苷酸测序引物，对含有酮还原酶域的这些PCR产物中的区域进行序，以证实所需序列已获得。
″179″切除体正向5′-CCTGATGGACGCGGGTGCGC(SEQ ID NO37)反向5′-GACACCGAAACCCCTG(SEQ ID NO38)″204″切除体正向5′-CCTGATGGACGCGGGTGCGC(SEQ ID NO39)反向5′-GCCGTGTGCACCACAGCGGTCAG(SEQ IDNO40)″260″，″283″，″306″切除体正向5′-(GTGTGATGTCGCCGACCGGCCCAGGTC(SEQ ID NO41)反向5′-GCGCTGGTGGGCCAGGGCGTCC(SEQ IDNO42)B.发酵产物的发酵和分析接种500μl的蓝灰链霉菌菌丝体片段的悬液(新鲜的或是冰冻的)于含有25ml的KB3培养基((10g/L细菌用胰蛋白胨，5g/L酵母提取液，3g/L牛肉提取液，1g/L KH2PO4，1g/L K2HPO4，1.5g/L Difco琼脂，pH6.8，浓度0.5ml/L含有30g/L FeSO4、30g/L ZnSO4.7H2O、4g/L MnSO4、4g/L CuCl2.5H2O和0.4g/LCoCl2.6H2O的一种痕量金属溶液)的种子烧瓶中。培养物在31℃下以220rpm振荡培养48小时。将500μl的种子培养物转到含有25ml的SD2生产培养基(85.5g/L葡萄糖，0.36g/L KCl，0.72g/L MgSO4.7H2O，7.2g/L CaCO3，4.86g/L(NH4)2SO4，0.72g/L K2HPO4，7.2g/L pharmamedia，浓度为1.8ml/L含有30g/LFeSO4、30g/L ZnSO4.7H2O、4g/L MnSO4、4g/L CuCl2.5H2O和0.4g/L CoCl2.6H2O的一种痕量金属溶液)的生产烧瓶中。培养物在31℃下培养10天。从第120小时(通常)开始，并且一直持续到发酵的结束，100μl等份的产物培养物被移出，并被加入至900μl甲醇。所得悬浮液震荡1分钟，并通过在微量离心管13,000rpm转速下离心10分钟，从而分离。10μl的提取液通过反相HPLC分析。
为了用反相HPLC分析试样，在装有Waters Model 996光电二极管排列探测器，Waters Model 717自动加样器以及Waters Nova-Pak C18柱(8mm×100mm)的Waters Model 625液体色谱仪上对样品进行色谱分析。该柱是用含有60％(v/v)乙腈，以及40％(v/v)100mM醋酸铵的流动相，在pH4.5的条件下，以2ml/min的流速平衡和洗脱的。目的化合物Fα和23-酮Fα(莫西菌素的前体)通过监测其在242nm的光吸收度，以及与真正样品相比较的保留时间而被检测。
在前述内容中，提供了本发明特定实施方案的详细描述，但其目的只是用于阐明本发明，而不是用于限定本发明。应该理解的是，基于所公开内容的，对于本领域技术人员显而易见的所有其它的修饰，衍生物以及等价物也是打算包含在本发明要求保护的范围内的。
序列表<110>Huang，ChengjinChaleff，DeborahRuppen，MarkStephens，Jerome<120>为抗生素的生物合成从蓝灰链霉菌非产蓝亚种中克隆基因及其使用方法<130>AM100484<140>60/471,256<141>2003-05-16<160>42<170>PatentIn version 3.2<210>1<211>88400<212>DNA<213>细菌<400>1gagctcttcg ctcccgccgg accggttggt cgcgccggag aggacgagcc ggtagcgggt 60gttgatctcg ttggtgccga gcccgttggc cgggcggccg tggaaccacc tcggatcggg 120ctcgggggtc tcctggccct tcttcagcgg cagatggtac ggctggccga tcagcgagga 180gccgacggcc ctgccgtccg ccgtgatctc ggagccgtcg gcccggtcgc ggaagagtgc 240ctgggcgacg ccggtgacga ccagcgggta gccggcgccc gtcaccaggg tcagcacgag 300gagggcccgc aggcccgccc cgagcagccg gacggtgtgg gtggcggagt tgttcatggc 360ggtcagcacg ctttcgtgac gtcacggccc gggaacgagg gagatgaaca ggtcgatgat 420cttgatgcct atgaagggcg ccaccaggcc gcccaggccg tagatcccga ggttgcgccg 480cagcatccgg tccgcgctca ccggccggta ccgcacgccc ctcagggaca gcggcaccag 540cgccacgatg accagcgcgt tgaagatcac cgcggagagg atcgcggagt cgggtgagga 600
caggcccatg acgtcgagtc gctccaggcc gggatgggcc ggcgcgaaca gcgccgggat 660gatcgcgaag tacttcgcga cgtcgttggc cagggagaag gtcgtcagtg cgccgcgtgt 720gatcagcagt tgcttgccga tctccacgat ctcgatcagt ttggtgggat cggagtcgag 780gtcgaccatg ttgccggcct ccttcgcggc cgacgtaccg gtgttcatcg ccacgccgac 840gtccgcctgg gccagagccg gggcgtcgtt ggtgccgtcc ccggtcatgg cgaccagcct 900gccgcctgcc tgctcccgcc tgatcagcgc catcttgtcc tcgggagtcg cctccgcgag 960gtagtcgtcg acgcccgcct cgcgcgcgac ggcctgcgcg gtcagcgggt tgtcacccgt1020gatcatgacg gtcctgatgc ccatgcggcg cagttcctcg aaccgcgcgc gcatgccgtc1080cttgacgacg tccttgaggt ggacggctcc cagcacccgg gcgccccgct cgtcccgcgc1140ggcgaccagc aggggcgtgc cccccgatcc ggcgatgcgg tcggcgatgg ccttcgcgtc1200ctgggcggcc tcaccgccct gctcctcgac ccaggcgagg atggaaccgg ccgcgccctt1260gcggatcctg cggccgccga cgtccacgcc cgacatgcgg gtccgggcgg tgaacgcgat1320ccattcggcg ccggcgagtt cgccccggtg ccgctcgcgc agtccgtact gctccttcgc1380caggacgacg acggaccggc cctcgggcgt ctcgtccgcg agcgaggaga gctgcgcggc1440gtccgccacc tcggcctccg tggtgccgga caccggcacg aacccggccg cccgccggtt1500gccgagcgtg atcgtgccgg tcttgtccag cagcagcgtg gagacgtcgc ccgcggcctc1560gaccgcccgg cccgacacgg ccagcacatt gcgctgcacc aggcggtcca tgcccgcgat1620gccgatcgcc gagagcagcg cgccgatcgt ggtcgggatg aggcagacca gcagcgccac1680cagcaccgtc ggtgtcaggt gggtgcccgc gtgatccgcg aagggcggca gcgtggcgca1740gaccagcagg aagacgatgg tcagcgaggc cagcaggatg ttcagcgcga tttcgttagg1800cgtcttctgc cgggccgcgc cttcgacgag gtcgatcatc cggtcgatga aggtctcacc1860gggcttggtc gtgatccgga tgacgacacg gtcggacagg accttggtgc cgccggtgac1920
ggcgctccgg tctccccccg actcgcggat gacgggtgcc gactcgccgg tgatggcgga1980ctcgtcgacg gacgcgacgc cctcgacgac atcaccgtcg ccggggatga cgtccccggc2040ctcgcagacc accagatcgc cgatcctcag tccggtgccc ggcacccgct cctccgagcc2100gtcctcgcgc aggcggcggg cgacggtgcc ggtcctggtc ttgcgcaggg tgtcggcctg2160tgccttgccg cggccttcgg cgaccgcctc cgcgaggttg gcgaagagca cggtcatcca2220gagccaggcg gagacggtcc agccgaaccg gtcgccggga tccatgaggg cgaagacggt2280ggtgaggacc gagccgatcc acaccacgaa catcacgggc gtcttgatct gcacccgcgg2340gtccagcttg cggaaggcgt ccggcaacga cctgacgagc tggcccgggt cgaacagacc2400gccgccgacc cgcctttcgg acggctggtg accggtgggg gcgtcgcgct gcggcgtccg2460ggcgggagtg atcgtggaca tcgggttccc ttggtcgtcc gggtgtgcgc tcatgccgcc2520agcccttcgg cgagcggccc cagcgccagg gccgggaagt acgtcaagcc ggcgaggatc2580aggatcgcgc ccaccatcag gccgctgaac agcggcttgt cggtgcgcag ggtgccggtg2640gtgaccggca cgggccgttg cccggcgagc gagccggcca gcgccaggac gaacaccatc2700ggcaggaagc ggccgagcag catcgccagt ccgatggtgg tgttgaacca ctgcgtgtcc2760gcgtcgagac cggcgaaggc cgagccgttg ttgttggcgc cggaggtgta ggcgtagagg2820atctcggaga acccgtgcgc gccgctgccg gtcgtcgagt tcaccggcgt cggcagggcc2880atcgcgcacg cggtgaggat caggaccagc gccggggtga ccagcaggtg gcaagcggcc2940agtttgatct cgcgggtgcc gatcttcttg cccaggtact cgggcgtgcg gccgaccatc3000agaccggcga tgaacaccgc tgtgacggcc atgacgagca tgccgtagag gccggatccc3060accccgcccg gagcgatctc gcccagcatc atgccgagca tcgcgatgcc tccgccaagg3120ccggtgaagg aggagtggaa ggagtccacc gcgccggtcg aggtgagcgt ggtcgacacc3180gcgaagatgg acgaggcgcc gacaccgaag cggacctcct tgccctccat cgcgccaccg3240
gcgatctcga gcgccgggcc gcggtgcgag aactcggtcc acatcatcag ggcgacgaag3300gcgatccaga aggtggccat cgtcgccagg atcgcgtagc cctgcctgac cgagccgacc3360atgacgccga acgtccgggt gatcgagaag gggatcacca ggatcaggaa gatctcgaag3420aggttggtga agggcgtcgg gttctcgaac gggtgggcgc tgttggcgtt gaaatagccg3480ccgccgttgg tgcccagcag tttgatggcc tcctgggagg cgaccgcgcc cccgttccac3540tgctgcgagc cgcccgtgaa ccggccgacc tcgtggatgc cggagaagtt ctggatgacc3600ccgcaggcgg ccagcaccac ggcgccgagg gtggccagcg gcaccaggac gcggaccgtt3660ccgcgcacca gatcggccca gaggtttccc agttcaccgg tgcgggagcg cgcgaacccg3720cgcaccagcg cgaccgcgac ggccatgccc acggcggccg aggtgaagtt ctgcacggcc3780aggccggcgg tctgcacgag gtgtcccatg gcctgttcgc cggagtacga ctgccagttg3840gtgttggtca cgaaggacac ggccgtgttg aacgcctggt ccgggtcgac ggctcgaaag3900ccgagggaca gcggcaggac gccttgcgcc cgctggacca ggtacaggaa gaggacgccg3960gccacggaga aggccagcac accgcggagg tacgcgggcc agcgcatctg ggcgccgggg4020tcgacaccga tgccccggta gatccatctc tcgacgcgcc agtgctcgtc ggaggagtag4080accttggcca tgtggttgcc gaggggtttg tggacgagtg ccagagcact cgtcagggcg4140agcagttgga gcacgccggc gagtacggga cccatggctg ctctcagaac ctctccggga4200agatcagggc gaggacgaga tagcccagca gggagacggc cacgaccagg ccgacgacgg4260tctcggcggt cacagcttcg tcacccccct ggcgacaaca gccaccagcg cgaagagcgc4320gagcgtggtg acgacgaagg ccgtatcggc catcgcggac tcctggaatg aggtgcggtg4380gaaacggacc cttgcaggta agcgcctcac cgaccgaaac aggacgtccg ttgacgtttc4440ccttacggcg tgacgtacgt ctttgacgga actcttacgc ctgaggtccg tgtccatgcc4500cctcggtccg ttgcggacca tgcccccgcg gccaccggag agggcggcgt ccccctcagc4560
gggccccgcc cgtctccccc ggggcctccg tctcccccac cttctccacg accgtctcct4620ccggcccgac cggccgtccg tccgcggcac ggatccgcaa ggggcgcagc gggcggccgg4680tgcggcggtc gagcaggcgg gagtggtcct cgccgggggc gaagcactgc tcctcgcccc4740actggcgcag ggcgacgatc accgggaaca aggcgcggcc cttgtccgtg aggacgtact4800cacggtggga accgccgtcc ggcgcgggca cgttgcgcag taccccggcc tcctccagcg4860cgcgcagccg cgccgtcagg atgttcttgg cgatgccgag gctgcgctgg aactcgccga4920agcggcgact gccgtcgaag gcgtcccgca cgatcagcag cgaccaccag tcgccgatgg4980cgttcaccga ccgggcgacg ggacaggggt cggcgtcgaa acgggtgcgg gcgaccatcc5040gcgtctctcc tctcctccgg caccccggat ccctccaggg atggttgcaa catgctacct5100cgtacggcta ccgtcctcgc cggtagcaag atgcaaccga gtgagaggtg tgacggtatg5160gcggtccagt gctccggtgc ggacggcgga tgcggcgaag ccggtggtcg cggagcggcc5220ggcacggcgc cgcccgcgcg gctcgtgccc ctgctcgccc tggcctgtgg cagctccgtc5280gccaccgtct acttcgccca ccccctgctg gtgaccctcg gtgagcgctt cgcgctcggc5340cccgggctgc tcggcgcgat cgtcaccgtg acgcaactcg gttacgcggt gggcctgctg5400acactcgtgc cgctcggcga cctgctcggc caccggcggc tggtcaccgc tcagctcgga5460ctgctggcac tggcgctgct ggccgccggg ctggcgccgg gcgcggctgc gctgctcggc5520gcgctcgccg cggtcgggct gctcgccgtc gtcgcccaga cgatggtcgc ggctgccgcc5580gccctgagcc cgcccgaccg gcgaggccgc gccgtgggaa ccgtcaccgg cggcatcgtc5640accggcatcc tgctggcgcg cgccgccgcg ggcgtcctcg ccgacctcgc cggctggcgg5700gcggtctacc tggcgtcggc gggcgtcacc gccgtcctcg ccgtgctgct gcgccgtgcg5760ctgcccccgg gatcgccgtc cgcaaaggct cgcgagacgt cgtacgtacg gctggtggcc5820tcgaccgtca ccctgttcgc ccgccatccg ctgctgcgga tccggggggc cctggccctg5880
ctggtgttcg cggccttcag cacgctgtgg agcggcgtgg cccagccctt gagcgatccg5940ccgtggtcgc tgtcgcacac cgcgatcggc gcgttcgggc tcgccggggc ggccggagcc6000gtcgccgcac aggtggccgg gcgctggaac gaccgggggc tcgcccggcg cacgaccggc6060gccggcctcg cgctgctggc gctctcctgg ctcccgatcg ccctgacccg gcaatcgctg6120tgggcgctgg cgatcggcgc cgtcctgctg gacttcgccg tgcaggccgt ccacgtcacc6180aaccagaccc tcatccacgc cgtccggccc gaggcgggca gcaggatcat cggtggttac6240atggtcttct actccgcggg cagcagcctc ggcgccctcg gttcctccct cgcctacgcc6300acggcgggct ggccggccgt gacggccctg ggcgcgtcgt tcagcgtcgc cgcgctgctg6360ctgtggacgg cgacccgtcg tacggggctg cccggcgacg acccggcggc cgaacggacg6420gaccctggcc gtccgtccgg ggacagggct gccgggaggc ccgcccgcag ccgctcttcc6480ggcccccggt gaacgtctgg ggtggcgcgg ggcgcgcgat aggggtccgc cgagagtcag6540gggttgtccc gcctgtacag cggcatcggc ggtcggtcca atggaaggcg tgcctccgga6600tgcggggcgg gtcgtcgacg accctgtgcc ggcggcggat tcccaccctc ccccggaagg6660cgaggtcgtg atgaccgacg tccgtcatga cagcaggcag acgggtccgg cgctgcgcgc6720gctcagcgcg gcccggcggg cgcgggcctt ggcgtccgcg atggcggcgg ccgccgcgga6780gacgcggcag gccgtcgagg ccgcggacgg cacggaccgc gcggccgccg tagccgagat6840cggcgcggta ctggaggacg cctcccggca cacggacgcc gccgccgagg ccgccgcctc6900ggctgccgag gccgccgccc gggccgagac ggccgaggcc gcccgcacgg tggccgccga6960gtcggccgag gccgtcgtcg ccgccgccga gacggcggtc cgggcggccc gggtcaccga7020ggccgccacg agcgccgccg cccaggccgc ggccggtacg gacgcggcgg gcgtgatggc7080ggacgccgcg gcgcacaccc ggcaggccac cgccgagacc gcggcgatcg ccgaggccgc7140cgccgcggcg gccagcgcgg cccgggccgc cgtcggcgac gaggcggcgg acggcgcgga7200
cccgtgccga cgggctgacg aggcggaggc cgcggccctg cggctgtgcg aggacacgcc7260gtggctgcgc aggcacctcc ccgacgtgtg aggcagggtg cccggcggcg ccggcgcgag7320atggaacccg ggccgggcgg ccctcttccc tccgggtgcc ggcgacgaga ccgtcgcccc7380cacctcgaac cgggccttcc ggtcgctgta ccggcatccg gagatcgagc ggcggccgca7440ctccgagggg gaccgggtgc tcggcggccg ggccgccacc tggacggatc cgccgtcgct7500ggagctgacg ggccgggtgg tccacgacgc gctgcgcctg ttccccggac gggctgctca7560ccgggtcatc accgaggaca cggggcgcgc agggcgcgcg ccgccggccg gcggcgtcgt7620cgcctgggtc gatcagcagc cccccgcggc gggttgctca cgttcggcgg cgggcaggcg7680tccgcccggt gcgcgttcag accgggcttt cgatgtgcgc gcacagccgg gcgggcagtt7740cctgacggcg gctgatctcc agcttgcggt aggcgcgcgt caggtgctgc tccacggtgc7800tgacggtgat gcagagccgg gcggcgatct cccggttggt gtgcccgttg gcggcgagtt7860ccacgaccct gagctccgat ccgctcagcg ccgcctcggt ccgccccgtg ccgtccccga7920acgactgccg cccaggacct cccggcagga tccgctcgca cagagcccgc gccccgcagt7980cgttcgccag gtgccaggcg cggcggatcg tggcgcccgc ccgggtcgac tcgccgcgct8040cccggtaggc ggcgcccaga tcggccaggg cacccgccag ggccagccgg tcgccgctgc8100tcttcaggtg gttcacggcc tcggtcagca ggttcagccg atccggcggt tcggcgatct8160gcgcgcgcag ccgcagcgag acgccgcgca catgaggatc gtcgtccggg gtccgggcca8220gttgttcccg caggagccga tcggccctcc tcggctcgca cagccgcagg aacgcctccg8280ccgcgtccga gcgccacggc atcagcgtcg gccggtcgat cccccagcgc cgcagcagac8340ggccggcgcc gaggaagtcg cggacggcgg cgaggggccg gtccagggcg aggtagtagt8400ggccccgggc gcgcaggtag gcggggccgt acacgctgcg gaacagggcc tccggcaccg8460ggtggtcgag ctgccgggtg gcctcgtcgt agcgccccat cgcggtggcc gcgaacaccc8520
ggctggccag cggcccgccg atgaagacgc tgcggctgca cctcggcacg caggccaggg8580cctgacgggc gtactcctcg gtgtcggcga gcagcccccg gcacagcgcg atctcggcct8640ggagggcgag gaactgcgcc ttccagcccg gcagccggcg accggcggcc tcgccgagca8700gacgtgtgca ccagagcgcc gcggtctcgt acgagccggt gcggcacagg acccggaccg8760cgttgacgac gatcaccagg gtggtgtcgg tgagcggcag gctcctcagg acgtgctccg8820ccgcgtcgga ggccgaggcg ttggtgccgt cgccggggag gtcccagatc ccggtcaccg8880gcatccgcgg gcgggggctc tcctcgtccc cgggctccgg gtccgtccgg ggacggatca8940gcggctccca gagcgcggag gcgtggaagc ccgtctccag ccggggagtt cgcgggtcgc9000cgtgggggcc cggccgtccc atgacctccg tggcctcctc cagccgtccg cagccgagca9060gcaggtgacc cagccgttcg gtctcggcgc tcgtcagtcg tccggcccgc agctcggtga9120cgagttcggc gaggtggtcc tccgccgccg ccgggtcggt gcgccgggtg gcgacggtca9180ggcgcagcag gatctcggcg cgccggggcc ctcccgcaca ggagcgccgg gccagttcga9240gacaggagac ggcggtcagg acgtcgtccc gcatcagcag ctgctcggcg gcgtcccgca9300gcacggacat cgcccagggc ccggccgcgt gccgggcggc gagcaggtgc cgggccacct9360cgtccggttc cgcgccgacg tcgtacagca gcgcggcggc gcggcggtgg aggtgtgcgc9420ggtggtcgtg gtccagggtg tccagggcgg ccgcctcgac cacggggtgc cggaagcggc9480cggacgccgt caggccggtc gcctccaggg cgcgcagccc gcgggccgcc atggcgcggc9540cgatgccgag cagccgggcg atcacctcgg cgcagccgga gtcaccgagg acggcgagcg9600cgccggcgct gtgcctaacc aggctgtcgg tgcgggacag tgaggcgagg acggcctggt9660agaaccgccc gccgatgacg ggcgacgctg ctctccggcg ccggccggca cgctcgtcgg9720tgtgcccttg ggtgtggctt tcgacgagtt cttcgagcag ggcatgcacc agcagcggat9780tgccgccggt gacggcgagg aggtcgtccg cgggcagggc ctcgacggcc ggtccggggc9840
gggcggcgcg cagtccggac acggcgcgca gggagaggcg gccgagcatg acgcgctgga9900gggccggctg gcacagcagc tcggcctcca cggcggggtc ggccgccagg ccggacggca9960gcgcggtgca gaccagcagc agtcgggtgg cgcggggatg gtcgacggcc tggagcaggc 10020agtgcaggga ctgcgggtcc gcgtggtgca ggtcgtcgat gccgatgacg accggcgcgg 10080cgccggtgag ctggtggagc gccgcccgca cacgctgcgc ggccggggtc tccgtcccga 10140cggcgtcctg gagcagtgag cgctgggcgt ccgggatgtc ggggtcgacc gccagttgcc 10200gcaggaggtc gaaggggcgc cggccctccg gcgggcttcc ggcggaccgg aggaccagga 10260agcccgatgc cgccgcgtgc ttgagcgcct cccccaggaa cgcgcttttc ccgcagccga 10320gtcctccttc gacgaccagc acccgcaccc ggccggcggc gcacgcctcg agcgccgttc 10380tcacggcatg ggactgcctg cccaggccga ggaacgtgag cccttgcggc tcccgcacgg 10440acaccgaagg ggaaacgccc cgcataatct ccctctgact ccctcccccg aagaccgggg 10500gctttacgga ttcgtaccaa caggaaagcc cacaagtcga cgagatactg cccctctccc 10560gaagccgcca cacgcgcacc ccgatacgag aatgagccaa tgagcaagcg tggtggccga 10620gttgatacga acccgtgaat ttacgttatt tcgctcaccc tttcgagcgt gtggagagtc 10680ctcggaatgg gcggccggga ggttgggcag cctccgcggg acggcgagcc attcgcgagg 10740tcacgcggac acgcgtgttg cgataatcgc acttaaggag aggacgagcg atgcccgacc 10800tttgcgagac cgaatccctc tggctccggc ggttccagcc ggctcccgcg gcccggacgc 10860ggctcatgtg cttcccgcac gcgggcgggt ccgccagcgc ctatctgcgc ctggcccggt 10920ccctcgcccc cggcatcgag gtcctggcgg tccagtaccc cggacgacag gaccggcgcg 10980ccgagccctg cccggactcc gtcgaaggcc tggcggacga tctgttcgcg gccgtccggc 11040accgcgtgga cgcgtcgacc gcgctgttcg gacacagcat gggcgcggtc ctcgccttcg 11100agctggcccg gcggctggag cgcgacgcgg gggtccgctg cgcccggatc ttcgcctcgg 11160
ggcgccgggc accctcccgg ttccgtgacg actccgcccc ggccgccagc gacgcctcga 11220tgctcgccga gatgcggact ctcggcggaa ccgacctgcg ggtgctccag gacgaggaac 11280tgctgatcgc cgcgctgccc gcgctgcgcg ccgactaccg cgcgatcggg acctaccgcg 11340ccgccgacga cgccgtggtc ggctgcccgg tcaccgtgct ggtcggtgac gccgatccga 11400ggaccagcct cgacgacgcc cacgcctgga gcgcccacac cacggcggag tccgaggtgc 11460tcaccttctc cggcgggcac ttcttcctcg acgcccacca cgacgcggtg gtggaggtcg 11520tcaccgcgcg cctgcggcag gaccgcgcgc cccggccgga ccgggtgtga gggggcccgg 11580cccgaagggc cgggccgctc cgcgcgtctg ccggcaccgg gccgcaccgg acccggcgcc 11640ggcagacgcg cggcgacctc acatcatggc gggcgccagg gccattcccc cgctggcgtc 11700cagcagttgg ccggtgatcc agcgggcgtc gtcggagacc aggaaggcga cgatgccggc 11760gatgtcgttc ggccggccca gccggccgag cgcggtcagg gccgagatgc ccgcctcggc 11820ccccggggtc tcgcgcaccc agcggttcat gtcggtgtcc gtgatgccgg gggccacggt 11880gttgacggtg atgccgcgcg aaccgagttc gttggcgagc cggggagcca tcatctccag 11940cgcccccttg gtcatggcgt agggcagcag cggccaggcg atccgggtga cggccgagga 12000gacattgacg atgcgtccgc cgtcggccat cagtgacagg gcccgctggg tcacgaagaa 12060cggtgcccgg acgttgatgc ggtacacgcg gtcgaactcc tcgggcgtgg tgtccgacag 12120gccggggaca tagccgtcct gtgccgcgag cgccgggtcg ccgggggcgg gggcgacggc 12180cgcgttgttc accaggatgt gcagcggacg cccctccagc tcccgctcca gtgcggtgaa 12240gagctcatcc acggcgtcgt cccggaggag gtccgcccgg accgcgaagg cccgtccccc 12300cgcgcgttcg atcgtctcca ccgtctcctg ggcgctcttt tcctgcgttc cgtagtgcac 12360ggcgacccgg acgccctcgg cggcgagtcg ctgggcgatg gcttttccga tgccgcgcga 12420ggcacccgtg accaaggccg tcctgtcgtt caattccggc atcccgaatc cccttctgcc 12480
gattatctta cttttcctct tgatgcatgg ggtcggaccc gaggccagat ccgcaccccg 12540gccacgcgtg aggtcgcgac ctcaccgatt actgtgccag agtccaggcg acacacggga 12600gggcgggaat gcgatcgatt tccgcacccg gaactcgtag ggggagcaag aagatcggcc 12660gaatacccct ggggtggata gggggtacca ggaccgtcgg gcgatcacta ttttgaaaca 12720cgactccggc gcgcggccgg cggcgaaagt cctctccatg ccgggctgtc ccctgcctcg 12780aaatacctgc ggcgactttc gccctgcgat gcggccgccc atccctgccg agcggtgagg 12840agacgacaag tgcacgagac acacgcgcac ggcgaggaag ggtcgtccga cgggtccgcg 12900gacgcagtgg tcttcgtctt ccccggacag gggtctcagt ggccggggat gggtgcggaa 12960ctgtgggaca cctccccggt gttccgcgag agtgtgcgcg cctgcgccga cgcgctcgcc 13020ccgtacctcg actggtccgt cgaaggcgtc ctgcgcggcg ccccggacgc cccggccggc 13080ccggcgctcg atcgcgccga cgtcgcgcag ccggccctgt tcaccctcat ggtgtcgctg 13140gccgagctct ggcgctcgca cggagtcgaa ccctgcgccg tcctcgggca cagcctcggc 13200gagatcgccg ccgcgcatgt ggccggcgcc ctgaccctgg ccgacgccgc ccgggtggcg 13260gccctgtgga gccgggccca ggccacgctg tcgggcaccg gcacccttct cgcggccaag 13320gccgcccccg aggaactggc accgcacctt cagcggtgga acggcgacga ccggcacggc 13380acccggctcg cgatcgccgg cgtcaacggg cccggcagca cggtggtggc gggggacctc 13440gacgcgatcg ccgcgctggc cgccgacctg gcctcggcgg gggtgcggac ccgccgggtc 13500gccgtcgacg tgcccaccca ctcccccgcg atgcggaccc tgcgggaacg gatcctcacc 13560gacctggcct ccgtcgcccc gtgcgtctcc cgtctcccct tccactcctc gctcaccggc 13620ggtctggtgg acacccgcgg gctggacgcc gactactggt accgcaacat cagcgagacc 13680gcgcgcttcg acctcgccgc ccgcggtctc ctggccgacg gacaccggac gttcgtggag 13740ctgagcccgc acccgatact caccctgggc ctgcaagcgc tcgccgacga cgtccccggc 13800
gccgccgacg cgctcgtgac gggcacgctg cgccgcgggc gcggcggaat gcggcagttc 13860caggacgcgc tcggccggct cagcgtcccc gcgggcgggc ggcccggccg cgaggtgagc 13920gccgcggccc tggccggccg gctggcgccg ctctccccgg cgcagcagga gcatctgctg 13980gtggaattgg tctgcgccca cttcgccgca ctcgtcggcg gcgacggcgg ggcgccgccg 14040acggtgcggc cgtcggccgc cttcaccgat cagggctgcg actccgccac cgccctggag 14100ctgcgcgacc ggctccgcga ggcgaccggg ctgcgcctgc ccgccacgct ggtcttcgac 14160cacccgacgc cggccgcggt cgccggccgg ttgcgccgac tcgccctcgg gatcgaggag 14220acggcggaca cggcaccggt cgccgtccgc ggccaccggg agggcgaacc gatcgcgatc 14280gtcgggatgg cctgccgctt cccgggaggt gtccggtcgc cggaggacct gtggcggctg 14340gtcaccgaag gcggtgacgc gctcgggccg ttccccaccg accgcggctg ggacaccggc 14400cgccacgcgg aggacccggc cacacccggc acctacgtcc agggcgaggg cggattcctg 14460tacgacgcgg gcgagttcga cgccgagttc ttcgggatct ccccgcgtga ggcgctggcc 14520atggacccgc agcagcggtt gctgctggag atggcgtggg agaccttcga acgggcggga 14580atcgatccca cctcggcccg gggatcgcgt accggcgtct tcgccggggt cctcccgctc 14640ggctacggcc cccgcatgga cgagacggac cagggcaccg ccgacctcca gggccatctc 14700ctcaccggca cactgcccag cgtcgcctcg ggccgcatct cctacaccct cggcctggag 14760ggcccggcgg tgtcggtgga gacggcctgc tcgtcgtcgc tcgtcgccct ccacctcgcc 14820tgccgctcgc tgcgggcggg cgagtgcgac ctcgccctga cggggggcgt ctcggtgctg 14880gccaccctcg gcctgttcgt cgagttctcc cggcagcgtg gactgtcggc ggacggccgg 14940tgcaaggcgt acgcggcggc ggccgacggg accggatgga gcgagggtgc cgggctgctg 15000ctggtcgaac ggctctccga cgcacggcgg ctggggcacc gggtgctcgc ggtggtccgg 15060ggcagcgcga tcaaccagga cggcgcgtcg aacgggctga ccgcccccag cgggccgtcc 15120
cagcagcggg tcatccgcga ggccctggcc gacgcgggcc tgacggcggc ggacgtcgac 15180gcggtggagg ggcacgggac cggcacacga ctgggcgacc cgatcgagat cgaggcgctg 15240ctcgccacct acggacaggg acgcgcccgg gaacggccgc tgtggctcgg atcgctgaag 15300tcgaacatcg gtcacaccat ggccgcggcg ggggtgggcg gggtcatcaa gatggtgatg 15360gcgctgcggc acggggagct gccccgcacc ctgcacgtgg acgcgccctc gccccgggcc 15420gactggtcgg cgggcgaggt acggctgctg acggaggccg tcgcgtggcc cgcggcggcg 15480gacggtgagc cgcggcgggc cggggtgtcg tccttcggcg tgagcggcac caacgcgcac 15540gccatcctgg aggaggcgcc cgccccggag gacgaggaac cggcgccgcc ggacggtgaa 15600gcactactgc cgtgggcggt gtccacgcgg tcggaggccg cactgcggac gcaggcacgg 15660atgctggcgg acgtcgtacg cgacgacccc ggagtcggac tcgccgatgt gggtgcggag 15720ctggcccggg ggcgggcggc tctcgagcac cgggccgtcg tcatcgcctc cgggcgcgcg 15780gagttcgcgc gggcgctgga ggcggtggcg tccggcgagc cgcacccggc cgtggtccgg 15840ggccacgcgg ggagcgagcg cggcggagtg gtgttcgtct tcccgggcca gggcggtcag 15900tgggccggca tgggactcga cctcctgcga agctcaccgg tgttcgcgga gcacatcgcg 15960gcctgcggca aagctctggc cccgtgggtg aagtggtcgc tcacggaggt gctgcaccgg 16020gacgccgagg atccggtctg ggaccgggcc gacgtcgtcc agccggtgct gttctcggtc 16080atgacgtcgc tggcggcgct gtggcgctcg tacggcgtcg agccggacgc cgtgaccggg 16140cactcgcagg gggagatcgc cgccgcgtac gtctgcggag cgctcggtct ggaggacgcc 16200gcacggacgg tggcgctgcg cagccgcgcc ctggtggcgc tgcgcgggcg gggcggcatg 16260gcgtccgtcg cctccgccgc cccggacgtc gaggagctca tcgcgcggcg ctggcccggc 16320cggctgtggg tcgcggcgtt caacggcccc ggcgcggtga ccgtttccgg ggacggtgat 16380gcgctggagg agttcctggg ccactgcgcg gacacggagg tgagggctcg gcgcgtcccg 16440
gtggactacg cctcccactg cccgcacacg gaggcgatcg agcgggaact gctcgacgcc 16500ctggaggaca tcaccccccg gccggcggcg gtcccgttct attcgacggt cgacgacgcg 16560tggctggaca ccacacggct ggacgcctcc tactggtacc gcaacctgcg ccggcccgtc 16620cgtttcagcc aggccgtgcg cgccctcacg gacggcggcc accgcgtctt catcgaggcg 16680agcccgcatc ccaccctcgt ccccgccatc gaggaccacg gcgacgtcac cgccctcggc 16740accctgcgcc gccacggcga cgacaccgag cggttcctca ccgccctcgc ccacctccat 16800gtcaccggag ccgccggcca ggacctctgg cgccaccact acgcccggct caggcccgcc 16860ccccgccacg tcgacctgcc cacctacgcc ttccagcgcg accggtactg gtggagcggc 16920ggcgccgggc gcggggacgt caccaccgcc ggtctgcacc ccggcggcca tcccctcctc 16980ggcgccgcgc tggacctcgc cgacggcggc ggccgcctcc acaccggccg tgtctccctg 17040cgcacccacc cctggatcgc cgaccacggc gtcgcgggca tcaccctcct gcccggcacc 17100gccttcctcg aactcgccct gcacacgggc gagtcgggga acgtgcggga actcaccctg 17160cacgcgcccc tggtcgttcc cgacgaggag ggcgtcgacc tgcaagtgca cctcgcccgg 17220cccgacgaag cgggcctgcg cgccctgacc cgtcttctcc cgggccgcgg ggtgccgacc 17280ccgagagccc cctggcagcc ccacgccacc ggccttctcg ggccggccga ccgagcaccc 17340ggctcctccg gcctcgagcc gcacgacctg ggcggcgcct ggcctccgcc gggggcggtc 17400cccctcgtcc ccggcgaact cggcgacgtg cccggctgct acgcccgcct ggccgacgag 17460gggttcgagt acgggccggc cttccggggg ctgcgtgcgg tgtggcgccg cggcacggag 17520atcttcgccg aggtcgccct cccggccggc gacggctccg tgttccggct gcatccggcg 17580ctgctggacg ccgtgctgca ccccgtcgta ctcgggctgg tggacggcgt gccggcccgt 17640ccgctgccct tctcctggaa cggcgtggcg ctgcacgccc ccgcgagcgg cgcgctgcgg 17700gtgcgcctcg cgccggccga cgacggcgct gtcggcatca cggccgcgac ggccgccggt 17760
gagccggtgc tctcggtcgc cgcgctggcc ctgcggtccg cctcggcgga gcagttgcgc 17820gcggcgatcc gctccgcggc gggctcgcgc gacgccctct acgagctgga ctggctgccg 1788Cctcccggcgg accgggccgc ttcgcccggt ggggccgaca tcgcggccct gggcacatcg 17940gagctgccct gccgtacgta cgagaccatc gcggagctgt cgcaggccct cgccgacggt 18000gctcccgccc ccgacgccgt cgtctccgac gtcggcgccg tcggcgggcc gctggacacc 18060gtgagcctgc acggcctctg ccggcgcggg ctggaactcg tgcaagcctg gctgggcgag 18120ccccggacgg ccgacacgcg gctggtgctc gtgacgcgtg gggcggtcgg ctgtgccccg 18180gccgagccgg tcgccgatcc ggccgcggcc gcgctgtggg ggctggtgcg gtccgcgcag 18240gcggagcacc ccggacggct gctcctgctg gacctcgacc ccgccgggtc gcggcccgtc 18300tccggccgcc tggtggaaca ggcggtggcc tgcggtgagc cgcacatcgc cgtacggggc 18360gacggcctgc gcgtcccccg gttgtcccgc gcgacggccg cccccgcaca ccctcccgcc 18420ggtggccggg aagcgcagtg ggacccggaa gggaccgtcc tcatcaccgg cggcaccgga 18480agtctcggcg cgctgttcgc ccggcatctg gtgaccgcgc acggggtacg gcggctgctc 18540ctcgccagcc gcagtggccc cggcgccccc ggcgccgccg ggctgcggga cgaactgacc 18600gctcacggag ccaccgtcac cgtcgccgcc tgtgatgtgg ccgaccggga ggccgtcgcc 18660gccctcctgg cgtccgtgcc gtccgagcac ccgctgaccg ccgtagtgca caccgccggc 18720gtgctggacg acggcgtact cgcctcgctc accgccgacc ggctggcccg cgtcctgcgt 18780gccaaggccg acgccgcgct ccacctgcac gatctcaccc gcgatctgcc gctcgccgcc 18840ttcgtcctct tctcctccgt cacggcgacg ctcggcacac ccggccaggc caactacacc 18900gccgccaacg cgttcctcga cgcgctcgcc cggcatcggc gcgccgcggg cctgcccgcc 18960gtctcactcg cctgggggct gtgggagcag accggcgggc tgaccgatca cctcggatcg 19020gtcgacctgc ggcggatggc ccgcaacggc ctggtcgcgc tgcccgccga cgccggcctg 19080
gcgctcttcg acaccgcgct ggccctggac cgcgccaacc tggtcccggc gcggctcgac 19140ctgcccgcgc tgcgccgcgc cacacacgtg ccgcccgttc tgcggcggct ggtcgaggtg 19200ccgggggcgc cgagcgcgga ccggtccgcc gggtccggcg gcgaggtgag gccgctgcgt 19260gagacgctgg ccgggctgga cgaccggaaa cgccccgctg ccgtctcccg cctggtccgc 19320aggcacgtcg cgtgggtgct cggcgccgac ggtccggagt cggtggacga ggaccgcagc 19380ttccgcgacc tcggcttcga ctcgctgatg gccgtcgaac tgcgcaacca gctcaacacc 19440gccgccggca tccggctcgc ggccaccctc gtcttcgacc acccgacacc gtcggccgtg 19500gcgcggcacc tcctcgaccg gtgctcgccg gacccggccg ccccggccgc tccctcgggt 19560acggcggtcg cgtcggcgct cgccactctg gccgagctgg agacggcttt gaacggcatc 19620ccggccgagg agtggacggc cgccgggggc ccggcccggc tgatgacgct ggcgtcctcg 19680ctgcccgcgc ccgcgtccgt ccctcggaca ccggcggccg gcgaagccgc cgagaagctc 19740gcccacgcct cgcgcgacga gatcttcgcg ttcatcgatc gggagctggg gcgtgactcc 19800gggccagcct caccctctcg cctcggtccg cagacccccg actcgacaga caaggcgccc 19860tttcatggag aatgaggaaa agctcctgga ctacctcaag tgggtcaccg ccgatctgca 19920ccgctcgcgg gaacgcgtca ccgagctgga ggaggccggc cgggagccga tcgccatcgt 19980cgggatggcc tgccggttcc cgggcgaggt gcggtcgccg gaggagctgt gggggctggt 20040cgcctcgggc ggcgacgcga tcggggcgtt cccggacgac cgcgggtggg atctggacgg 20100gctgttcgac cccgacccgg agcgtgcggg cacctcgtac acccggcgcg gcggtttcct 20160gtacgacgcg gcggagttcg acgcgggctt cttcgggatc tccccgcgtg aggcgatggc 20220gatggacccg cagcagcggc tgctgctgga gacctcgtgg gaggctttcg agcgggccgg 20280catcgacccg tcctcggtac gcgggtcccg ggtcggtgtc ttcgccggcc tcatgtacca 20340cgactacgcg gcggcccagg gcagcaccgg ggacggagac ggggagccgg acttcgaggg 20400
ctacctcggc gacggcagcg tcagcagcat cgcctcgggc cgtatcgcct acaccctcgg 20460gctcgcgggc gcggcgatca ccgtcgacac ggcctgctcc tcttccctgg tcgccctgca 20520cctcgcctgc caggcgctgc gcaccggcga ctccgagctg gccctggccg gcggggtcag 20580cgtcatgtcc accccccgca ccttcgtcca gttctcgcgg cagcggggcc tgtcggcgga 20640cggccggtgc aaggcgtacg cggcggcggc cgacgggacg gggttctccg agggcgtcgg 20700catggtgctg gtcgaacggc tctccgacgc ccggcggctg gggcatccgg tactggcggt 20760cgtgcggggc agcgcggtca accaggacgg cgcgtcgaac ggtctgacgg cgcccaacgg 20820accgtcgcag gagagggtga tccgcgaggc gctggccaac gcgggcctga cggcggcgga 20880cgtcgacgcg gtggaggggc acgggaccgg gacacggctg ggtgacccga tcgagttgca 20940ggcgctgctc gccacctacg gacagggacg cgcccgggag cggccgctgt ggctcggatc 21000ggtgaagtcc aacatcggtc acgcgcaggc ggcggcgggg gtgggcggcg tcatcaagat 21060ggtgatggcg ctgcggcacg gggagctgcc gcgcaccctg cacgtggacg cgccctcgcc 21120ccgggtcgac tggtcggcgg gcgaggtacg gctgctgacg gaggccgtcg cgtggcccgc 21180ggcggcggac ggtgagccgc ggcgggccgg ggtgtcgtcc ttcggggtga gcggcaccaa 21240cgcccatgtg atcctggagg aggcgcccgc gtcggagggc gaggaagctc cgccgccgga 21300gcccgggtcg ccgttgccgt gggtggtgtc cggtcactcg gaggcgggct tgcgcgccca 21360ggcgcaggct ctggcggagt tcgcacggac cgcgcccggg gccgaactcg tggacgtggg 21420agcggcgttg gcccgggggc gggcggcgct ggggcatcgg gcggtcgtcg tcgcctcgga 21480gcgtgaggag ttcgagcggg cgctggccgc gctggcctgt ggcgaaccgc acccgtgtgt 21540ggtcgacggg tcggcggacg gccggcgcga ggacggtgtg gtgttcgtct tcccgggcca 21600gggcggtcag tgggccggca tgggactcga tctgctgacg acctcggggg tgttcgccga 21660acatatcggt gcgtgtgaac gcgcgctggc gccgtgggtg gagtggtcgc tgacggagat 21720
gctccaccgc gaggcggagg acccggtgtg ggagcgggcg gacatcgtcc agccggtgct 21780gttctcggtc atggtgtccc tggccgcgct gtggcggtcc tacggcatcg aacccgacgc 21840ggtggtcggc cactcccagg gcgagatcgc cgccgcccac gtctgcggcg ccctcaccct 21900cgaagacgcc gcgaaagtcg tggcactgcg cagccgggcc ctggccgcac tgcggggccg 21960cggcggcatg gtctccctct cgctgtcgac cgcggatgcc ggggagctgg tggagcggcg 22020gtgggccggg cggctgtggg tcgcggcgct caacgggccg gaggcgacga cggtctcggg 22080ggacgtcgac gcgctggagg agctcctggc ccactgcgcg aaaagcgagg tgcgagcgcg 22140gcgcgtcccg gtggactacg cctcccactg cccgcacacg gaagcgatcg cggaagagat 22200cgtcgattca ctcggggaca tcacgccccg ggccgccacc gttccgttct actcgacggt 22260cgacgacatg tggttggaca ccacacggct ggacgcctcc tactggtacc gcaacctgcg 22320cctcccggtc cgcttcagcc aggccgtgcg cgccctcacg gaagaaggcc accgcctctt 22380catcgagacg agcccgcatc ccaccctcgt ccccgccatc gaggaccacg gcgacgtcac 22440cgccctcggg accctgcgcc gccacggcga cgacaccgag cggttcctca ccgccctcgc 22500ccacctccat gtcaccggag ccgccggcca ggacctctgg cgccaccact acgccaggct 22560caggcccgcc ccccgccacg tcgacctgcc cacctacccc ttccaacgcc ggcgctactg 22620gctggagaaa cccgacccgc agaccaggcc ccagcggtcc cgctccaccg ccccggacct 22680cgacaggctg gaggcggagt tctggcaggc cgtcgaggaa accgacaccg acaccctcgc 22740ccacaccctc cacctcgaca cccagaccct cgaacccgtc ctccccgccc tcgccacctg 22800gcaccaacaa caacgcgacc acgcccgcat caacacctgg acctaccagg aaacctggaa 22860accactccac ctccccacca cccgacccac cacccccacc agctggctca tcgccatccc 22920cgaaacccac cgcaaccacc cccacaccac caacctcctc accaacctcc cccaccacaa 22980catcaccccc atccccctca ccatcaacca caccaccgac ctccaccacg cctaccacca 23040
cgcccaccac cacaccaccc cacccatcac cgccgtcctc tccctcctcg ccctcgacga 23100aacaccccac ccccaccacc cccacacccc caccggcacc ctcctcaacc tcaccctcac 23160ccaaacccac acccaaaccc acccaccaac ccccctctgg tacctcacca cccaagccac 23220caccacccac cccaacgacc ccctcaccca ccccacccaa gcccaaacca tcggactcgc 23280ccgcaccacc cacctcgaac acccccacca caccggcgga cacatcgacc tccccaccac 23340accccacccc aacaccctca cccaactcat caccgccctc acccaccccc accaccaaca 23400caacctcacc atccgcaccc acaccaccca cacccgacga ctcaccccca ccaccctcca 23460acccaccacc cccacaccac ccaccaaccc ccacggcacc accctcatca ccggcggcac 23520cggcgccctc gccaccaccc tcgcccacca cctcgccacc accggcaccc aacacctcct 23580cctcaccagc cgacgcggcc cccacacccc cggcgcccga caactccaca cccaactcac 23640ccaactcggc accaacacca ccatcaccgc ctgcgacctc tccgaccccg accaactcac 23700ccacctcctc acccacatcc cccccgaaca ccccctcacc accgtcatcc acaccgccgg 23760catcctcgac gacgccaccc tcaccaacct cacccccacc caactcgaca acgtcctgcg 23820cgccaaagcc cacaccgccc acctcctcca ccacgccacc ctccacaccc ccctcgacca 23880cttcgtcctc tactcctccg ccgccgccac cctcggcgcc cccggccaag ccaactacgc 23940agccgccaac gcctacctcg acgccctcgc ccaccaccgc cacacccaca acctccccgc 24000caccaccatc gcctggggaa cctggcaagg aaacggcctc gccgactcgg acaaggcccg 24060cgccaacctc gaccgccggg gcttcctgcc catgcccgag acgctggccg cagccgcggc 24120cgtgcgggcg atcgagagca ggcggccgtc cgtggtcatc gccgccatcg actgggccag 24180agccgagcgc acccccgacg tcgaggatct cctccccgcg gccgacgagg ggtcgtcgag 24240tggcaagccg gaggccgcgc cggtggacct gcgcggtacc ttgagccggc agtccgccgc 24300cgaccaacag gccacactgc tcggcctggt gcggacccag gcagccgtcg tactgcgcca 24360
cacggagccc gaggcgctcg ccccgggcca ggccttccgg gcgctcggct tcgactccct 24420caccgccgtc gaactccgca accgactggc caaggccacg gacctcgcgc tgcccgcctc 24480actggtcttc gatcacccga ctccggtgaa gctcgcggag ttcctgcgca ccgagctgct 24540cggcaccgca ccagctacca ccgccgccgt cccggccctc caggcacaca ccgacgaacc 24600catcgccatc atcggcatgg cctgccgctt ccccggcgcc gtcaccacac ccgaacacct 24660gtggaacctc atcgccaccg aacaagacgc catcggcgag ttccccaccg accgcggctg 24720ggacctggac aacctctacc accccgaccc cgaccacccc ggcaccacct acacccgcca 24780cggcggattc ctccacgacg ccggcgactt cgacgccgac ttcttcggca tcaacccacg 24840cgaagccctc gccatggacc cccaacaacg actcctcctc gaaaccgcct gggaagccat 24900cgaacacgcc ggcatcctcc ccgacgccct gcacggcacc cccaccggcg tcttcaccgg 24960cgtcaacgcc caggactacg ccgcacacac ccacacctcc ccccacacca ccgagggcta 25020caccctcacc ggaaccgccg gcagcatcgc ctccggccgc atcgcctacg tcctcggact 25080cgaaggcccc gccgtcacca tcgacaccgc ctgctcctcc tccctcgtcg ccctccacct 25140cgcctgccag gccctgcgag caggcgaatg caccacagcc ctcgccagcg gcatcagcat 25200catgaccaca ccgctggcct tcaccgagtt ctcccggcag cggggtctgg cggcggacgg 25260ccggtgcaag gcgttcgcgg cggccgccga cggtaccggc tggtcggagg gggtggggac 25320gctgctgttg gagcggttgt cggacgccga gcggaacggg caccgggttc tggcggtggt 25380gcggggcagc gcggtcaacc aggacggcgc ctccaacggg ctgacggcgc cgaacggtcc 25440gtcccagcag cgtgtgatcc gccaggccct ggtcaacgcg aacctctccg cagttgatgt 25500cgacgccgtc gaagcccacg gcacggggac caagctgggc gacccgatcg aagcccaggc 25560cctgctcgcc acctacggcc agggacgtgc gcaggaacag ccactgtggc tcggttcggt 25620caaatccaac ctgggtcaca cccaggcggc ggcaggcatg gccggcctga tcaagatggt 25680
gatggcgctg cggcacgagt cgttgccgcg gacgttgcat gtggatgagc cgtcgccgga 25740ggtggactgg tcgtcggggg cggtgagtct gctgaccgag gcgcggccct ggccgcgggt 25800cgaggaccgg ccccggcggg ccggggtgtc ctcgttcggg gtgagcggga cgaacgccca 25860cgtcatcgtg gaggaggcgc ccgcgccgac gggagtggag gcggtggaag ccgcgccggc 25920gggggtggag actgcggcgg ctgcggcggt ggtggtggag acggacggtg cgggccgggt 25980gtcggcggat ctgccgttgg tgtgggtggc gtcgggcaag tcgcaggccg cgatacgcgc 26040ccaagccgcc gccctgcacg cccacgtcct ggaccacccc gaacaggacg cggacgacat 26100cggctacagc ctggccacca cccgcgccct gttcgaccac cgcgccaccc tcatcgcccc 26160cgaccgccac accgtcccgg agcccctcac cgggctgggc gacggacgca cgcaccccca 26220cctcatcccc acacccccca ccgaacccgg ccacacccac aaaatcgcct tcctctgctc 26280cggacaaggc acccaacgcc ccggcatggc caccggcctc taccacacct accccgcctt 26340cgccgccgcc ctcgacgaaa cctgcgccca cttcgacccc cacctcgacc accccctgca 26400cgacctcctc ctcaaccacg accccaccga cctcctcacc cacaccctct acgcccagcc 26460cgccctcttc accctccaaa aagccctcca ccacctcatc accgaaacct acggcatcac 26520cccccactac ctcgccggac actccctcgg cgaaatcacc gccgcccacc tcgccggcat 26580cctcaccctc cccgacgcca cccacctcat caccacccgc gcccgcctca tgcaaaccat 26640gccccccggc accatgacca ccctccacac cacccccgaa cacatccaac ccctcctcga 26700ccaacacccc ggcaaagccg ccatcgccgc cgtcaacagc ccccactccc tcgtcatcag 26760cggcgacccc gacaccatcc accacatcac caccacctgc cacaaccaag gcatcaccac 26820caaacccctc gccaccaacc acgccttcca ctccccccac accgacacca tcctcgaaca 26880actcgacacc accacccaca ccctcaccta ccaccaaccc cacacccccc tcatcaccag 26940cacccccggc gaccccctca ccccccacta ctggacccac cagacccgcc aacccgtcca 27000
ctggaccgac accatccaca ccctccacac ccacggcgtg accacgtaca tcgcactcgg 27060accagagcac accctcacca ccctcaccca ccacaacgtc ccccaccacc aacccaccgc 27120catcaccctc acccaccccc accacaaccc cacccaccac ctcctcaccg cactcgccca 27180cctccacaca acccaaccca ccggccccaa catctggcac caccactaca ccccagtcgc 27240acccgccccc cgccacgtcg acctgcccac ctaccccttc ccacgccggc gctactgggt 27300gcaggcgtcc gccggtacgg gtgacgtgtc ggctgccggg ctccagcgac cggaccaccc 27360actgctcggc gcggtgatgg agctcgcgga cggggacgga atcgtcctca ccgggcgctt 27420gtccctgcac acccacccct ggctcgccga ccacagcgtc ggcggcgtcg ccctccttcc 27480cggtaccgct ctgctggagc tggcttttca ggctggtctg cgtgcgggtt gtcctggtgt 27540cgatgagctg actctccatg ctcctctggt ggttccggag tcggggcatg tggtggtgca 27600ggtgtcggtt tcggtgccgg gcgaggcggg tcgtcgtggt gtgagtgtgt acgggcggct 27660ggtggaggac ggggggctgg agggtgagtg gacgcggcat gccgagggtg tggtgtgtcc 27720gtctgttcct ggggagtcgg tggttgtgga gccggtggcg gacggggtgt ggccgccgtc 27780cggtgcgcag ccggtggatc ttgaggagtt ctacggtcgt ctggcgggtg ggggttttgt 27840ctacggtccg gtgttccagg gtttgtgtgc ggcctggcgg gacggggacg acgtggtggc 27900cgaggtgcgt ctgccggacg aggggctggc cgatgtcgcg ggcttcgggg tgcatccggc 27960gctcctggac gcggccgtgc aggcagtcac cctcctgttc ccggaccagc agcaagccgg 28020tctcgcggcc cacacatgga acggtgtctc gctccacgcc cggggcgcca ccgtcctgcg 28080cctgcgcatg actcccaccg acgcgacctc gaccgccgtt cgcctgcacg ccaccgacga 28140gaccggagca cccgttctca ccctcgactc gctcctgatg cgtccggtgc cgttggaggg 28200gctgggggcg ggggtgcggc gtggctcgtt gttcgagctg gggtgggtgc cggtggaggg 28260gatgccggcc tcggtggccg gtgggggcgg ggagttggtg gcgtgggagt gcccgggtgg 28320
tggggtggcc gaggtcacgg ccgcggcgtt gggagtggtg caggagtggc tcgccgatga 28380gcgggagggg gacgcgcggc tggtcgtggt gacgcgtggt gcggtcgcgg tggatgcggg 28440ggagccggtg cgggacgtgg cgggggccgc tgtgtggggg ctggtccgct cggcccagtc 28500cgagcatccc gaccggttcg ccctgctcga cctcgacccc gacaccaaga ccgaccccgg 28560catcgacacc gacggggaca ccgacgtgtc cgccgacgcg aaggtcggca ccggtgatgg 28620tctcgacgat gccgccgtcg cgtccgctct ggcccgcggt gagagccaac tcgccgtacg 28680cgacggggtg gttcgcgtag cgcggttggg gggtttggtt ggggggttgt cgttgcctgg 28740tggggtgggg tggcggctgg atggtggtgg gtcggggttg ttggaggggg tgggtgtggt 28800tgcttcggat gcggctgggg tggtgctggg tcgggggcag gtgcgggtgg cggtgcgggc 28860tgccggggtg aacttccggg atgttctggt ggcgttgggg atggtgccgg gtcaggtggg 28920ggtgggcagt gagggtgcgg gggtggtggt ggaggtgggg cccggggtgg agggcctggt 28980ggtgggggac cgggtgttcg gggtgttcgg ggacgcgttc gcgccggtgg tggtggcgca 29040ggaggtgttg ctggcccgta tcccggaggg ctggtcgttc gcgcaggcgg cttcggtgcc 29100ggtggtgttc gctaccgctt acctgggact ggtcgatctg gcgggggtgc ggcgggggga 29160gagtgtgctg gtccatgcgg cggccggcgg ggtcggtacc gccgcggtgc agctcgcccg 29220tcatctgggg gcggaggtgt atgcgacggc cagtgaggcg aagtgggcgc gtctgcgggc 29280ggcgggtgtc gcgccgcagc ggatcgcgtc ctcgcggagt gtggagttcg agtcccgttt 29340ccgccgggcc agtggcggcc ggggtgtgga tgtggtgctg aactgtctgg cgggtgagta 29400caccgatgcc tcgttgcggc tgtgttcgcc gcaggggggc cggttcctgg agctgggcaa 29460gaccgacatc cgtgatgccg gtgaggtcgc cgctcggttc ccgggggtgt cctaccgggc 29520gtatgacctg atggacgcgg gtgcgcagcg ggtgggggag atcctgcaca cggtggtgga 29580tctgttccgg cgcggggtgc tggagccgtt gccggtcacc gcgtgggacg tgcgccaggc 29640
ccatcaggca ctgcggtcga tgcggtcggg cctgcacgtc ggcaagaacg tgctcaccct 29700gcccgtgccc ctggatgcgg aggggacggt gctggtgacg ggcgggaccg gcactctggg 29760ggcggcggtc gcgcgccatc tggccgccgg gcacggggtg cggcatctgc tgctggtgag 29820ccggcgcggc atggccgccg ccggtgccga aaaactgtgt gcggaactgg gtcaggcagg 29880ggtttcggtg tcggtggccg ggtgtgatgt cgccgaccgc gcccaggtcg ccgccctgct 29940ggagcaggtg cccgcggagc atccgctgac cgctgtggtg cacacggccg gtgtcctgga 30000cgacgccacc gtgacctgcc tggaccggaa caagatcgat gcggtgctcg gggcgaaggt 30060ggacggtgcc ctgcacctgc acgagctgac cgcggggatg gacctgtcgg cgttcgtgct 30l20gttctcctcc gccgccgggg tcctgggctc gccggggcag ggcaactacg ccgccgccaa 30180cgccgccctg gacgccctgg cccaccagcg ccgcgccgcc ggtctgcccg ccctctccct 30240ggcctgggga ctgtgggaag aggccagcgg gatgaccggc catctggatg ccgctgaccg 30300tcaccgcatc acccgctcgg ggctgcatcc cctgaccacc cccgacgccc tcgccctcct 30360cgacaccgcc ctggccgccg gacgccccgc actcctgccc gccgacctac gccccaccca 30420ccccgcaccg cccctcctgg aacacctcgc gcccgcccgc accagccacc gcaccgcaca 30480caccagcacc gcaaccggcg tgggccagga cgtctccctc accgaccgcc tcgccaccct 30540gacccccgaa cagcggcacg acaccctgct ggcgctggcc cgtacccaca tcgccgccgt 30600cctgggccac cccagccccg acaccatcga ccccgaacgc accttccgcg acctcggctt 30660cgactccctc accgccgtcg aactccgcaa ccggctcacc cgcgccaccg gcctgcgcct 30720gcccgccacc ctcgccttcg accaccccac ccccaccgca ctcacccacc acctcaccac 30780cctcctcaac cccaacgaca acgacaacgt cggtccggta ctgatggagc tcgaaagact 30840ggaatccgct ctcgccgcgc tggacaggga cgacagcgcc tgcgagcggg tcactctgcg 30900actgcaatcg ctgatgctca ggtggagcgg ctccgagcgg cagtcagccg aaaacacgga 30960
cgactccagc aggttcgcgt cggcgaccgc ggaggagcta ctcgaattca tcgaccgaga 31020cctgggtctt tcctgaacca gctcggtctt ccctgaacca gctcgacgac gcggttttcc 31080cgtgcgcgac ggactccaag gacgtgaacc agacgtggcg aatgacgaga aggtgctcga 31140atacctcaag cgagtcaccg cggatttgga ccggaccagg cggcgcctgt acgaagtcgt 31200cgagcgggag caggagccga tcgccatcgt ggggatggct tgccgttatc cgggcggggc 31260cgggtcgccc gcaggtctct gggacctcgt cagctccggt acggacgcca tcggggagtt 31320ccccaccgat cgtggctggg atctggaacg tctctacgac cccgaccccg atcacccggg 31380caccacgtac acccgccacg gcggattcct cgacggcgta ggtgagttcg acgcggagtt 31440cttcggcgtc agcccgcgtg aggccctggc gatggacccc cagcagcggc tcctcctcga 31500aaccgcctgg gaagccatcg aacacgccgg catcgtcccc gagtcgctgc gcggcacgtc 31560caccggcgtc ttcgccggta tcaacccgca ggactacacc atcagtcagt acggacggga 31620ttcggagatc gagggctatc tgctgaccgg ggcagccgcc agtatcgcct ccggccgtat 31680ctcctacacc ctcggcctcg aaggcccagc cgtcaccatc gacaccgcct gctcctcctc 31740cctcgtcgcc ctccacctgg cttgccaagc gctgcgcgca ggggagtgca ccatggccct 31800ggcgggcggc gcctcggtcc tgtccacacc gctgatcttc gtcgagttcg ctcgccatca 31860cggcctgtcg gtcgacggcc ggtgcaaggc gttctccgct tcggccgacg gcacgggctg 31920gggcgagggc gccggcctgc tcctcctcga acggctctcc gacgccaagc gcaacggccg 31980ccgcatcctc gctctcgtac gggggagcgc ggtcaaccag gacggcgcct cgaacgggct 32040gacggcgccg aacggaccct cccagtgcag ggtcatccgc cgggccttgg ccaacgccca 32100tctcgccccg gccgacatcg atgccgtgga agctcacggc accggcacca ccctgggcga 32160ccccatcgaa gcccaggccc tccaggaagc gtacggcgcg gaccgacccg acgatcggcc 32220gctctgggtc ggcacgctca agtcgaacat cggccactcg atcgccgcgg cgggtgtggg 32280
cggggtcatc aagatggtga tggcgctgcg gcacgagtcg ttgccgcgga ccttgcatgt 32340ggatgagccg tcgccgcagg tggactggtc gtcgggtgcg gtgagtctgc tgaccgaagc 32400gcggccctgg ccgcgggacg aggaccggcc ccggcgggcc ggggtgtcct cgttcggggt 32460gagcgggacc aacgcgcacg tgatcctgga ggaagcgccc gcgccggcgg aggtgcaggc 32520ggtagaaact gcgccggtgg tgcgggtgga tggtggggag cgttccgcac cggcggatgt 32580gccgttggtg tgggtcgtgt cgggcaagtc gcaggccgcg ctacgcgccc aggccgccgc 32640cctgcacgcc cacgtcctgg accaccccga acaggacgcg gccgacatcg gctacagcct 32700ggccaccacc cgcgccctgt tcgaccaccg cgccaccctc atcgcccccg accgcgacac 32760cctcctggac gccctcaccg ccctggccga cggccgcacc cacccccacc tcgtccccgc 32820accccccacc gaacccggcc acgcccacaa aatcgccttc ctctgctccg gacagggcac 32880ccaacgcccc ggcatggcca ccggcctcta ccacacctac cccgccttcg ccgccgccct 32940cgacgaaacc tgcgcccact tcgaccccca cctcgaccac cccctgcgcg acctcctcct 33000caaccacgac cccaccggcc tcctcaccca caccctctac gcccagcccg ccctcttcac 33060cctccaaaaa gccctccacc acctcatcac cgaaacctac ggcatcaccc cccactacct 33120cgccggacac tccctcggcg aaatcaccgc cgcccacctc gccggcatcc tcaccctccc 33180cgacgccacc cacctcatca ccacccgcgc ccgcctcatg caaaccatgc cccccggcac 33240catgaccacc ctccacacca cccccgaaca catccaaccc ctcctcgacc aacaccccgg 33300caaagccacc atcgccgccg tcaacagccc ccactccctc gtcatcagcg gcgaccccga 33360caccatccac cacatcacca ccacctgcca cacccaaggc atcaccacca aacccctcac 33420caccaaccac gccttccact ccccccacac cgacaccatc ctcgaacaac tcgacaccac 33480cacccacacc ctcacctacc acccacccca cacccccctc atcaccagca cccccggcga 33540ccccctcacc ccccactact ggacccacca gacccgccaa cccgtccact ggaccgacac 33600
catccacacc ctccacacca acggcgtcac cacctacatc gaactcggac ccgaccacac 33660cctcaccacc ctcacccacc acaacctccc ccaccaccaa cccaccgcca tcaccctcac 33720ccacccccac cacaacccca cccaccacct cctcaccgca ctcgcccaca cccccaccac 33780ctggcacacc caccaccaca cccacaccaa cccccacccc cacaccatcc ccgacctccc 33840cacctacccc ttccaacgcc ggcactactg gctccaggcg cccaccacca gcaccgatca 33900gccggtggcc ccgacgaacg acgacgcccc cgcgcctcga gcgacatcgc tccgggacac 33960tcttgccgga cgaagccctc aagagcgcga agaagtgctc ctggatctcg tactgaccca 34020ggtcgccgcc gtgctcggcc acaccgcgcc tgaggtggtg gatccccaaa gggcgttcaa 34080ggacctcggc ttcgactcac tggccgccat caaactccgc aacaggctcg ccgcagccac 34140cggactcgag ctgccgacca cccttgtctt cgaccacccc acgccggtgg cactccgcca 34200gtacttccag tcgcagatcc tcggagcgga ggcggacgcc cccaaccgtc tgcccctccg 34260ggcggcgacc accgacgaac ccatcgcgat cgtcggcatg gcgtgccgct tcccgggcgg 34320cgttcggacg gccgacgacc tgtggcagct cctgagcgac gaacacgatg cggtcggcgg 34380cttccccacc aaccggggtt gggacgtggc gaacctctac gacccggacc cggatcgcca 34440cggcaccacg tacacccagc agggcggctt cctctacgaa gcgggggagt tcgacgccga 34500gttcttcggc atcagcccgc gtgaggccct ggcgatggac ccccagcagc ggctcctcct 34560cgaaaccgcc tgggaagcca tcgaacacgc cggcatcaac cccgatgccc tgcgcaacac 34620gtccaccggt gttttcgccg gggtcatcta ccacgactac gcgagccggt tcctcaccgc 34680gccggccggt tacgagggct acctcggcca cgggagtgcc ggcagcatcg cgtcgggccg 34740tgtcgcgtac gtgctgggtc tcgagggtcc cgcggtcacg gtcgacaccg cgtgttcgtc 34800gtcgctcgtc gcgctgcatc tggcctgtca ggcactgcgg tcgggcgagt gcacgatggc 34860tctggcgggc ggcgcgacgg tgatgtcgac cccgcaggcg ttcgtggagt tctcccggca 34920
gcggggtctg gcggcggacg gccggtgcaa ggcgttctcc gctgcggccg acggcacggg 34980ctggggcgag ggcgccggcc tgcttctcct cgaacggctc tccgaggccg agcggaacgg 35040acaccgggtt ctggcggtgg tgcggggcag cgcggtcaac caggacggcg cctcgaacgg 35100gctgacggcg ccgaacggtc cgtcccagca gcgcgtgatc cgccaagctt tggccaactc 35160gggcctgacc ggcgccgatg tcgacgccgt cgaagcccac ggcacgggga ccaagctggg 35220cgacccgatc gaagcccagg ccctgctcgc cacctacggc caggaacacc accccgacca 35280gccgctctgg ctcggctccc tgaagtccaa catcggccac gcccaagcgg cagcaggcgt 35340gggcagcatc atcaagatga tcatggctat gcgcaacgag tcgctgccgc ggacgttgca 35400cgtggatgag ccgtcacccc atgtggactg gtcgtcgggg gcggtgagtc tgctgaccga 35460gccacgcccc tggccacgcc gggaagaccg gccccggcga gcgggaatct cctccttcgg 35520agtcagcggg acgaacgccc acgtcatcgt ggaggagccg cctgcgcggg cggaggtgga 35580ggcggtggaa gccgcgccgg cgggggtgga gactgcggcg gctgccgcgg tggtggtgga 35640gacagacggt gcgggccggg tgtcctccga tgtgccgttg gtgtgggtgg tgtccggcaa 35700gtcgcaggcc gcgctacgcg cccaggccgc cgccctgcac gcccacgtcc tggaccaccc 35760cgaacaggac gcggccgaca tcggctacag cctggccacc acccgcgccc tgttcgacca 35820ccgcgccacc ctcatcgccc ccgaccgcga caccctcctg gacgccctca ccgccctggc 35880cgacggccgc acccaccccc acctcatccc cacacccccc accgaacccg gccacaccca 35940caaaatcgcc ttcctctgct ccggacaagg cacccaacgc cccggcatgg ccaccggcct 36000ctaccacacc taccccgcct tcgccgccgc cctcgacgaa acctgcgccc acttcgaccc 36060ccacctcgac caccccctgc gcgacctcct cctcaaccac gaccccaccg acctcctcac 36120ccacaccctc tacgcccagc ccgccctctt caccctccaa aaagccctcc accacctcat 36180caccgaaacc tacggcatca ccccccacta cctcgccgga cactccctcg gcgaaatcac 36240
cgccgcccac ctcgccggca tcctcaccct ccccgacgcc acccacctca tcaccacccg 36300cgcccgcctc atgcaaacca tgccccccgg caccatgacc accctccaca ccacccccga 36360acacatccaa cccctcctcg accaacaccc cggcaaagcc accatcgccg ccgtcaacag 36420cccccactcc ctcgtcatca gcggcgaccc cgacaccatc caccacatca ccaccacctg 36480ccacaaccaa ggcatcacca ccaaacccct caccaccaac cacgccttcc actcccccca 36540caccaacacc atcctcgaac aactcgacac caccacccac accctcacct accacccacc 36600ccacaccccc ctcatcacca gcacccccgg caaccccctc accccccact actggaccca 36660ccagacccgc caacccgtcc actgggcgga caccatccac accctccaca ccaacggcgt 36720caccacctac atcggactcg gacccgacca caccctctcc accctcaccc accacaacct 36780cccccaacac caacccaccg ccatcaccct cacccacccc caccacaacc ccacccacca 36840cctcctcacc gcactcgccc acacccccac cacctggcac acccaccacc acacccacac 36900caacccccac ccccacacca tccccgacct ccccacctac cccttccaac gccggcacta 36960ctggctggag gtcccgaagc cgactgccga agcatccgcc tcagccagtg gcccggggcg 37020gaaccgggcc gccaaactct cagcgctcga ggcggagttc tggcaggccg tcgaggaaac 37080cgacaccgac accctcgccc acaccctcga cctcgacacc cagaccctcg aacccgtcct 37140ccccgccctc gccacctggc accaacaaca acgcgaccac gcccgcatca acacctggac 37200ctaccaggaa acctggaaac cactccacct ccccaccacc cgacccacca cccccaccag 37260ctggctcatc gccatccccg aaacccaccg caaccacccc cacaccacca acctcctcac 37320caacctcccc caccacaaca tcacccccat ccccctcacc atcaaccaca ccaccgacct 37380ccaccacgcc taccaccacg cccaccacca caccacccca cccatcaccg ccgtcctctc 37440cctcctcgcc ctcgacgaaa caccccaccc ccaccacccc cacaccccca ccggcaccct 37500cctcaacctc accctcaccc aaacccacac ccaaacccac ccaccaaccc ccctctggta 37560
cctcaccacc caagccacca ccacccaccc caacgacccc ctcacccacc ccacccaagc 37620ccaaaccatc ggactcgccc gcaccaccca cctcgaacac ccccaccaca ccggcggaca 37680catcgacctc cccaccacac cccaccccaa caccctcacc caactcatca ccgccctcac 37740ccacccccac caccaacaca acctcaccat ccgcacccac accacccaca cccgacgact 37800cacccccacc accctccaac ccaccacccc cacaccaccc accaaccccc acggcaccac 37860cctcatcacc ggcggcaccg gcgccctcgc caccaccctc gcccaccacc tcgccaccac 37920cggcacccaa cacctcctcc tcaccagccg acgcggcccc cacacccccg gcgcccgaca 37980actccacacc caactcaccc aactcggcac caacaccacc atcaccgcct gcgacctctc 38040cgaccccgac caactcaccc acctcctcac ccacatcccc cccgaacacc ccctcaccac 38100cgtcatccac accgccggca tcctcgacga cgccaccctc accaacctca cccccaccca 38160actcgacaac gtcctgcgcg ccaaagccca caccgcccac ctcctccacc acgccaccct 38220ccacaccccc ctcgaccact tcgtcctcta ctcctccgcc gccgccaccc tcggcgcccc 38280cggccaagcc aactacgcag ccgccaacgc ctacctcgac gccctcgccc accaccgcca 38340cacccacaac ctccccgcca ccaccatcgc ctggggaacc tggcaaggaa acggcctcgc 38400gagcggtgac atcggcgagc atctgcgccg ccgcgggatg atcccgctgg atcccgagtc 38460cgctgtcggt gccttcgacc gggcggtcgc gagcgatcgg cccagcgtct tcgtcgcgga 38520catcgactgg cccaccttcg gccgcaacac ctccagcggt cttcgcgccc tcttcgagga 38580cattccggag gccacacagc ctgagccgac cgcccggagc gcggaccagc cgaacgggca 38640cggtagcctc caggaacttc tcgcccgcca gtccccggcc gagcaggccg aaacgctcct 38700ggcattggtc cggacgcatt ccgcgaccgt cctcgggcgt gacggggccg atgccgtcgc 38760cgccgaacgt cccttcaggg acctgggatt cgactcactg tccgccgtcg agctccgcaa 38820tcatctgacg gccgacacgg agctcgctct gccgacaacg ctggtcttcg atcacccgac 38880
tccggtgaag ctcgcggagt tcctgcgcac cgagctgctc ggcaccgcac cagccaccac 38940cgccgccgtc ccggccctcc agtcccacac cgacgaaccc atcgccatca tcggcatggc 39000ctgccgcttc cccggcgccg tcaccacacc cgaacacctg tggaacctca tcgccaccga 39060acaagacgcc atcggcgagt tccccaccga ccgcggctgg gacctggaca acctctacca 39120ccccgacccc gaccaccccg gcaccaccta cacccgccac ggtggtttcc tctacgacgc 39180cggcgacttc gacgccgagt tcttcggcat caacccacgc gaagccctcg ccatggaccc 39240ccagcaacga ctcctcctgg aaaccgcctg ggaagccatc gaacacgccg gcatcctccc 39300cgacgccctg cacggcaccc ccaccggcgt cttcaccggc gtcaacgccc aggactacgc 39360cgcacacacc cacgcctccc cccacaccac cgagggctac accctcaccg gaaccgccgg 39420cagcatcgcc tccggccgca tcgcctacac cctcggactc gaaggccccg ccgtcaccat 39480cgacaccgcc tgctcctcct ccctcgtcgc cctccacctc gcctgccagg ccctgcgagc 39540aggcgaatgc accacagccc tcgccagcgg catcaccgtc atgaccagcc cggtcacgtt 39600caccgagttc tcccggcagc gagggctcgc ccccgacgga cactgcaagg cgttctccgc 39660ctcggccgac ggcaccggct ggagcgaggg cgtgggcacc atcctcgtcg aacggctctc 39720cgacgccgag cggaacgggc accggattct ggcggtggtg cggggcagcg cggtcaacca 39780ggacggcgcc tccaacggcc tgacggcgcc gaacggcccc tcccagcaac gcgtcatccg 39840ccaggccctg gccaactccg gcctgaccgg cgccgatgtc gacgccgtcg aagcccacgg 39900cacgggaacc aaactcggcg accccatcga agcccaggcc ctgctcgcca cctacggcca 39960gggacgtgcg caggaacagc cactgtggct cggctcggtc aaatccaacc tcggccacac 40020ccaggcagcg gcaggcatgg ccggcctgat caagatggtg atggcgctgc ggcacgagtc 40080gttgccgcgg acgttgcatg tggatgagcc gtcgccgcag gtggactggt cgtcgggtgc 40140ggtcagcctg ctgaccgagg cgcggccctg gccacgccgg gaggaccggc cccggcgagc 40200
gggaatctcg tccttcgggg tgagcgggac gaacgcgcac gtgatcctgg aggaggcgcc 40260cgcgccggcg gaggcggtgg agacggaaca gggtgtggtg ccgcagggcg accaggagtg 40320ttccgcgccg gtgggtgtgc cgttggtgtg ggtggtgtcc ggcaagtcgc aggccgcgct 40380acgcgcccag gccgccgccc tgcacgccca cgtcctggac caccccgaac aggacgcggc 40440cgacatcggc tacagcctgg ccaccacccg cgccctgttc gaccaccgcg ccaccctcat 40500cgcccccgac cgcgacaccc tcctggacgc cctcaccgcc ctggccgacg gccgcaccca 40560cccccacctc atccccacac cccccaccga acccggccac acccacaaaa tcgccttcct 40620ctgctccgga caaggcaccc aacgccccgg catggccacc ggcctctacc acacctaccc 40680cgccttcgcc gccgccctcg acgaaacctg cgcccacttc gacccccacc tcgaccaccc 40740cctgcgcgac ctcctcctca accacgaccc caccgacctc ctcacccaca ccctctacgc 40800ccaacccgcc ctcttcaccc tccaaaaagc cctccaccac ctcatcaccg aaacctacgg 40860catcaccccc cactacctcg ccggacactc cctcggcgaa atcaccgccg cccacctcgc 40920cggcatcctc accctccccg acgccaccca cctcatcacc acccgcgccc gcctcatgca 40980aaccatgccc cccggcacca tgaccaccct ccacaccacc cccgaacaca tccaacccct 41040cctcgaccaa caccccggca aagccaccat cgccgccgtc aacagccccc actccctcgt 41100catcagcggc gaccccgaca ccatccacca catcaccacc acctgccaca cccaaggcat 41160caccaccaaa cccctcacca ccaaccacgc cttccactcc ccccacaccg acaccatcct 41220cgaacaactc gacaccacca cccacaccct cacctaccac caaccccaca cccccctcat 41280caccagcacc cccggcgacc ccctcacccc ccactactgg acccaccaga cccgccaacc 41340cgtccactgg gcggacacca tccacaccct ccacaccaac ggcgtcacca cctacatcgg 41400actcggaccc gaccacaccc tctccaccct cacccaccac aacctccccc aacaccaacc 41460caccgccatc accctcaccc acccccacca caaccccacc caccacctcc tcaccgcact 41520
cgcccacacc cccaccacct ggcacaccca ccaccacacc cacaccaacc cccaccccca 41580caccatcccc gacctcccca cctacccctt ccaacgccgg cactactggc tggaggtccc 41640gaagccgact gccgaagcat ccgcctcagc cagtggcccg gggcggaacc gggccgccaa 41700actctcagcg ctcgaggcgg agttctggca ggccgtcgag gaaaccgaca ccgacaccct 41760cgcccacacc ctcgacctcg acacccagac cctcgaaccc gtcctccccg ccctcgccac 41820ctggcaccaa caacaacgcg accacgcccg catcaacacc tggacctacc aggaaacctg 41880gaaaccactc cacctcccca ccacccgacc caccaccccc accagctggc tcatcgccat 41940ccccgaaacc caccgcaacc acccccacac caccaacctc ctcaccaacc tcccccacca 42000caacatcacc cccatccccc tcaccatcaa ccacaccacc gacctccacc acgcctacca 42060ccacgcccac caccacacca ccccacccat caccgccgtc ctctccctcc tcgccctcga 42120cgaaacaccc cacccccacc acccccacac ccccaccggc accctcctca acctcaccct 42180cacccaaacc cacacccaaa cccacccacc aacccccctc tggtacctca ccacccaagc 42240caccaccacc caccccaacg accccctcac ccaccccacc caagcccaaa ccatcggact 42300cgcccgcacc acccacctcg aacaccccca ccacaccggc ggacacatcg acctccccac 42360cacaccccac cccaacaccc tcacccaact catcaccgcc ctcacccacc cccaccacca 42420acacaacctc accatccgca cccacaccac ccacacccga cgactcaccc ccaccaccct 42480ccaacccacc acccccacac cacccaccaa cccccacggc accaccctca tcaccggcgg 42540caccggcgcc ctcgccacca ccctcgccca ccacctcgcc accaccggca cccaacacct 42600cctcctcacc agccgacgcg gcccccacac ccccggcgcc cgacaactcc acacccaact 42660cacccaactc ggcaccaaca ccaccatcac cgcctgcgac ctctccgacc ccgaccaact 42720cacccacatc ctcacccaca tcccccccga acaccccctc accaccgtca tccacaccgc 42780cggcgtcaac cattacgctc ccgtggcggc gaccgacccg tccacgttcg cgtccgtcct 42840
cgccgcgaag gcggccggcg cggcacacct gcatgaactc ctgctggagc tggacacggt 42900cgagcagttc atcctcttct cctccggttc gggggcctgg ggcagcggca accagtgcgc 42960gtacgcggct gccaacgcct acctcgatgc gctggcggcg caccgccagg cccgcggcct 43020gcctggcatg tcgctcgcct gggggccttg ggacggtgac gggatgtcgg ccggagagga 43080cgcccagcgg tacctccgtg agcggggcgt actgcccatg gatccgcggc tcgccgtcgc 43140ggccttcgac gaggcggtcc gggcgcggcc gaactccaac ctcgtcgtcg cggacatcga 43200ctgggagcgt ttcgtcccga cgttcaccgc gcggggccac aaccccctga tcgaggacat 43260ccccgaagtc cgccggctgg ccgcggaggc cgaggccgcc cagaccacga ccgccgccac 43320ggacgccccc gcccttctca accgactctc aggtctgtcg gccactcagc agaagcagca 43380tcttctccgg ctggtgcggt cacacatggg cgaggtcctc ggccgcgagg acgtcgacac 43440gctcgacgag cgccacacct tccgggacct gggcttcgac tcgctcacct cggcccgatt 43500cagccagcgg ctcgccaagg acacggggct gcaccttcct gccaccctcg tcttcgacca 43560cccgacgccc gccgactgcg tggctcatct gcgggatcaa cttctgggtg aaacggacga 43620catgactccg aggaagcgag atcacctcgg ggaggaccgg cgggcggcca ccgcggacga 43680cccgatcgcg atcgtcggga tggcgtgccg gttcccgggc ggcgtgcggt ccgccgatga 43740tctgtgggac ctgctgtcgt cgggcaccga cgccatcagc ggcttcccca ccgatcgcgg 43800ctgggacatc gagagcctct acgaccccga ccccgaccgc tccggcacca cgtacacccg 43860ccacggtggt ttcctctacg acgccgggca gttcgacgcc gagttcttcg gcatcagccc 43920gcgtgaggcc ctggccatgg atccccagca gcggctcctt ctcgaaaccg cctgggaggc 43980cgtcgaacac gcaggcatca acccgcagac actccacggc acccccaccg gcgtcttcac 44040gggcgtcaac gcccaggact acgcagccca cctgcgccag gcgtcgggca acgtcgaggg 44100gtacgccctg accggaagct cgggcagtgt cgtgtcgggt cgggtggctt acaccttcgg 44160
tttcgagggg ccggccgtct cggtcgacac cgcgtgctcg tcgtcgctcg tcgcactgca 44220cctcgcaggc caagccctgc ggtccggcga gtgcacgatg gccctcgccg gcggcgtcat 44280ggtgatgtcc tcccctgaga cgttcgtgga gttctcgcgg cagcggggtt tgtcggtgga 44340cgggcggtgc aagtccttcg cggccgcggc cgacggtacc ggctggggcg agggcgtggg 44400catgctgctc gtggagcggt tgtcggacgc cgagcgcaac gggcaccggg ttctggcggt 44460ggtgcggggc agcgcggtca accaggacgg cgcctccaac ggcctgaccg caccgaacgg 44520cccctcccag cagcgcgtga tccgccaggc cctggccaac tccggcctga ccggcgccga 44580tgtcgacgcc gtcgaagccc acggcacagg aaccaaactc ggcgacccca tcgaagccca 44640ggccctgctc gccacctacg gccaggaaca ccaccccgac cagccgctct ggctcggctc 44700cctgaagtcc aacatcggcc acgcccaagc agcggcaggt gtcggcggga tcatcaaaat 44760ggtgatggca ctgcgccacg agacgctgcc gcgcacgctg cacatcgacg agccgacccc 44820ccaggtcgac tggtcgtccg gcgcggtcag cctgctgacc gagccccgcc cctggccacg 44880ccagggggac cggccccgac gcgccggcat ctcctccttc ggagtcagcg gaaccaacgc 44940ccacgtcatc ctggaagagg cacccgccca gccggccggg gaccccgccc cagaagacgg 45000cgccccggtg ccctgggcga tgtcggcgcg ttcaaacgcc gcgctgcggg cacaggccgc 45060actcctgcgt gacttcctcc aaggccccgg caccgacacc gcactacggg cggtcggagc 45120cgaactcgcc catggcaggg ccgtcctgga acaccgcgcc gtgatcgtgg cacgggaacg 45180gacagagttc gaagacgcgc tggaagcact ggcctcgggt gaaccgcacc ccgcactcat 45240cgaagacacg accggcagcc agaccaacag ccactccggt ggcggggtgg tgttcgtctt 45300ccccggccag ggcggtcagt gggccggcat gggactcgac ctgctgcgcg actcccaggt 45360gttcgccgac catgtcggtg cgtgtgaacg cgcgctggcg ccgtgggtgg agtggtcgct 45420caccgaaatg ctccaccggg acgcggagga tccggtgtgg gagcgggcgg atgtggtcca 45480
gccggtgctg ttctcggtca tggtgtccct ggcggcgctg tggcggtcct acggcatcga 45540acccgaagcg gtggtcggcc actcccaggg cgagatcgcc gccgcccacg tctgcggcgc 45600actcaccctg gaggacgccg cgaagatcgt ggcactgcgc agccgggccc tggccgcgct 45660gcggggccac ggcggcatgg cctcactcgc cctgaccgga accgaggccg aggacctcat 45720caccacccac tggccaggac ggctgtggac ggccgcgttc aacgggccac gggccaccac 45780cgtctccggc gacaccgacg ccctggacga actcctcacc cactgcaccg aaaccggggt 45840acgggcccgc cgcatccccg tggactacgc atcccactgc ccccacaccg aaaccatcga 45900acacgacctg ctccacatgc tccacggcat caccccccag cccggcagca tcccgttcta 45960ctccaccgtc gaggacgcct ggaccgacac caccaccctg gacgccgcct actggtaccg 46020caacctgcgc cggcccgtcc gcttcaccca cgccgtccgc accctcaccg cccagggcca 46080ccgcctcttc atcgagacca gcccccaccc caccctgacc cccgccatcg aagaccacga 46140ccacaccacc gccctgggca ccctgcgccg ccacgacaac gacacccacc gcttcctcac 46200cgccctcgcc cacgcccaca ccaccggcca caccgtcacc tggaccaccc actaccccac 46260caccccccac acccccgcca tcgacctgcc cacctacccc ttccaacacc accactactg 46320gctccacaca cccaccacca gcaccggcga cgtctccgcc gccggactgc accccaccga 46380gcaccctctc ctcggcgcca ccgtggaact cgccgacgga gacggaacct tgctcaccgg 46440gcgcctgtcc ctgcacaccc acccctggct cgccgaccac agcgtcggcg gcatcgtcct 46500cctccccggc accgccctcc tcgaactcgc cctcgaagcc gggacgcgca ccggttgccc 46560ccacgtccag gaactcaccc tgcacacgcc cctggtgatt cccgagaccg gacacgtcgt 46620cttccagctg acggtctcgg caccggacga gaccgggcag cgcccgttca ccgtccattt 46680ccgttccgag gccgtcaccg gcgcggacga tccggcggac cggacctgga cgcggtgcgc 46740caccggtgcg ctctcgaccg cggccgcccc cgatcactcc gaagccgcca cctggccgcc 46800
gccgtccgct cagccgctgg acctcgacgg tctgtacgac cgcatggcgg aggcgggtct 46860ggtctacggt ccggtgttcc aggggctccg cgaggcttgg ctcgatggcg aggacatcgt 46920cgccgaggtg cgcctgccgc aggaggcggc cgccgacacg cagggcttcg gcctgcatcc 46980cgccctgctc gacgccgctc tgcatgtgac ggcgctgacc tcacaggccg gtacagcgga 47040cgaagacgcg caggaacggc gtcggttgcc gttcgcgtgg gccggtgtct ccctgttcgc 47100cagggagtgc gcggcgctgc gtgtgcgggt ggcgccgtgt gcgccgcacc cgggggacgc 47160cgtggcgatc acagccaccg acgaggacgg ccgtccggtg ctggcggtgg aatcgctcac 47220cctccggccc gtctcccccg accagttgcg ggcggcggcc ccggccgccg ggcgggattc 47280gctgttccgc ctggagtggg taccggtcac ggcctccgcc tccgcctccg cccggccgac 47340cgggccctgg gccgccatcg gcaccggtcc ggcggtggcc ggcctggccg gccacgcaga 47400cctgacggtg tacgcggagg ccggcgatct gctccgggat ctggacggag gggcccccgc 47460gcccgctgtg gtcgtgctca gcgtcacgcc cgatgccgac gaattcgcca ctccccgtgc 47520ggcgaccggc cgggccctct ccgtccttca ggcctggctg gcggacgagc gcctggccga 47580cagccggctc gtggccgtca cttctggggc ggtcgtcgcc gcgcccgggg acgacacggt 47640cgacgtcccg ggtgccgccg tgtggggctt ggtgcgttcc gggcagtccg agcacccgga 47700ccgcatcacg ctgctcgact gtgcgagcgg cgcccggccc gggccggacc tcgtcgccgc 47760cgccctcgcc tcgggcgagc cgcagctcgc cgcccgcgcc ggggtcctct acacgccccg 47820gctggccagg ccgcaccgcg acgcctcggc cgtaccgcgg tcgctgccgt cccacggcac 47880cgtgctcatc accggcggca ccggtctgct gggcgggttg gtcgcccggc gcctggtgga 47940ggcgcacggt gtccgccgcc ttctcctggc cggccgcagg ggtccggcgg cggaggggct 48000ggactcgctg acgtccgagt tgcgtgagcg cggggcgacc gtcgaggtcg ccgcgtgcga 48060cgcggccgac cgcacacagt tggaggcgct gctggccggg gtgcccgagg agcatcccct 48120
gtccgcggtc gtgcacgccg cgggtgtgct cgacgacggg gttctcacgt ccctgacgaa 48180cgagcggctg ggagctgtcc tgcgggcgaa ggcggattcg gcgctgcttc tgcacgagct 48240cactcaggac ctcgacctgt ccgccttcgt cctgttctcc tccgccgccg gcgtcctcgg 48300ctctcccggc cagggcagct acgccgccgc caacgccgtg ctcgacgcac tcgcccacca 48360gcgcagcgcc gccggtctgc ccgctctctc cctggcctgg gggctgtggg cggagggcag 48420cgggatgacc gggcacctcg acgccgacga ccgctcccgg atcaaccggg ccggtatggc 48480gccgctcccg acgcccgatg ccctggatct gttcgacgcc gcgctgtcgt cggacgaacc 48540cttcctggta ccggctcgct tcgacctttc cgccgtacgc accaggaccg cgtacggccc 48600gctcccgccg ctgctgcgcg gcctggtccg gacctcgggc gcgcaccggg tccggggcgc 48660agtcggcgaa gcccgggcgg ccggcgtgga cgaggccgga cggctgcggg aacggctggc 48720ccgccagagt gacgccgaac gccggaacac cttgctgcgg ctcgtgcagt cgaacgtcgc 48780ggcggtgctc ggtcaccgcg gcacggggac cgtcgccgag acacgcgcct tccgtgagct 48840gggcttcgac tcgctcacgg cggtggagct gcggaaccgg ctgaaggtcg ccacagggct 48900ggcgctgcgg gccacggtcg ccttcgactt cccgactccg gcggcgctgg ccgagcatct 48960gggtgcccgc ctgcttccgc cggacggcgc cgtgtccgag gcggtgggcg agaaggagct 49020gcgcgggctc ttgacgtcga tcccgatcgg ccggctgcgg gaggcggggc tgatcgaccg 49080cctcctggcg ctcgccgctg cggcgccaga ctccgccgat cagacggcgg agcagccctc 49140ccggtccgtg tcggtcgagg acatcgacgc catggacgtc gacagcctca tcggcctggc 49200ccacgacacc ggcaccgact ccggtcacgc cccctgcgag ggctgacctc cacttcacgg 49260atgcgagaga cgacatgacg cagattccgc caaccggtca cgacgccgtg gcagccgggc 49320ccgcccccgg cgccgcggaa cagaaacgag gacggaaacg gaaaccagga cgggagcccc 49380ggccagagca tcgacgggaa caggaacgag ggcagggagc agggctgggg caggggcagg 49440
aacgcgcgcg gcccgcggac ggtggtcggc ggctcgtgct tggctgggcg gcgctcggcg 49500cggtgtgcct ggccctgcag gcgtacgtgc tcgtccgctg ggcggccgac ggtgggtatc 49560gcctggtgga cgtacccggt gagggcggcg cggagcgtgg ccaccgaagg gtcctcgaca 49620tcgtgttccc ggcgctgtcg gcggcaggtg tcgtggggct ggcgctgtgg ctctaccgca 49680ggtgccgcgc ggagcggcgg gtgtcgttcg acgccctgct gttcgccgga gtgctgttcg 49740cgggctggct gagcccgctg atgaactggt tccatcccgt cctggtctcc aacacgcacg 49800tgtggggcgc ggtcggctcc tgggggccgt acacgccggg atggcagggg tccgcccccg 49860ggatggaggc cgagctgccg ctggtgacgt tcagcgtgtg ctcgacagcg ctcctgggtg 49920tgctggcctg ctgtcacgtg ctgtcccgcg tccgggaccg gtggcccggg gtccgcccgt 49980ggcaactgat cggggtggcc gtcgccaccg cggtggccct ggacctgtcg gagccggcga 50040tctccttgat cggtctagtc cgtctggtcg aaggcgctgc cggaggtgtc gctgtggagc 50100ggtgcctggt accagttccc tctgtaccag ctcctgaccg cggccctggc cagcgggttg 50160ctgagcgcgc tccggttctt ccgcgacgag cgggacgaga cgctggtgga gcgcggtgcc 50220tggcgcctgc cgggccgtgt ccgcctctgg gcgcggttcc tggccgtcgt cggcggcgtc 50280catgtcgtga tgggcggcta tacggccctt catgtgctgc tctcgttggt cggcggccaa 50340ccgccggacg cgttgccggg gttcttccgt ccgccggccg tctactgagg gcggggcgga 50400cggcacgcaa cgaggggagg ggccggcgtc tcatgctctg ctgtccggtc agacctcagc 50460gcgctggcac ggcgcggtca ggacgacgta cccgatgtcc tccgtgtacc actggctgca 50520cttgcccacg aacgtctcca ggtcgtccgc cgtcatctcc agggcttcgg cgtaggcgtg 50580ggcgttggcg cgtacgtcgt cacccatcgc gctgtacgac ggggcgatga cgtgctcacc 50640gatgtcggtg agctcggtca gccggagtcc ggcgtcgctg atcatcccgg cataggcggt 50700gatggggatc agcgagggga cggcgagttc gctggacgac cagtccgccc ccgtcggctg 50760
tgatgcgcgg agtgtgacgt ccatggccgc cagccgccca ccggggcgca gcacacgggc 50820catctcctgg aacacccggg ccgggtcggg catgtgcagc aggcactcga gggcccagac 50880ggcgtcgaag gaggcgtcgg ggaagggcag gtccatggcg tcggcgcact cgaagcggac 50940ccggttcgcg agtccggacc gctcggcgag cgcggtggcc agctcgacct gccgggggct 51000gatggtgatg ccgacgatgt ccaccggctc gctgtgcgcc aggcgcaggg ccggccggcc 51060ggaaccgcag ccgacgtcca gcacacgtct gaccgggcgc ccggtgtgtt cccgaagctt 51120gccgatcata tggtcggtga ggcggtcgga ggcctggccg agtgtgctgc cgtcgtccgg 51180gtgcggccag tatccgaggt gcgtgttgcc gcccagggcc cggttcaaca ggctggtcat 51240gcggtcgtag tagtcaccga cgtccgcggg ggtcggtgat ccctggtgag gcgccttggt 51300catggttccg gcagctcctt cggtcgtgcg gcggcctcaa gggaggcgtc cgcgggggcg 51360tggccgcgag ggatggcggg ggtcctgggc tcggctatca tccgcaggcg gtcggggaag 51420acgtgggtcg ccttggcgac cgggcggacg cggtcgccct tgaggggacg cagacgccag 51480cgcgaggcga tgaccgcgac ggcgacggcc gtctccatga gggcgaagtt gtcgccgatg 51540cacttgtagg tgccgagcgc gaagggaacc caggcgccct tcggaacgtc gcgcgtggtc 51600tctttcgact cccagcggtc ggggtcgagc ttctccggat cacggtacca gcgggggtca 51660cgctggagcg cgtacgagct gtacatgatt tccacgtcgg ccggcagctc gtgttccccg 51720agccggacgg ggcgcaccgt gcgccgcgag cccacccagc cggggtactt gcgcagcgcc 51780tccttgacca ggcgctgggt gtacgggagg cgcgggaggt ccgcgctggt ggggagccgg 51840cctccgagga cggtgtcgat ttcggcgtgc agcctctgtt cgatgaggtg gtcgtgagcg 51900agttcgtgga agatccacgc ggtgagagcg gccggcccac cgattccggc gaccgcgagc 51960cccatgatct cgttgtgcac ctcgtcgtcc gtcatggtgt tgccctcggc gtcccgcgcg 52020cgcagcatcg tcgagagcag gtcgccgtgg tcgcggccgt cggcgcggta ggcggtgacc 52080
gcctcccgga tggcggcgct ggtgcggccc atgtggcgct tggcggcagt gggcagggag 52140gtgtagagct gcggggcgag cgcgctcagc ctggccacct tcaggatgtc gtgccccgtg 52200gtgcgcagtt ccgcctcggc cgccgcaccc aggtcggact ggaacaacgc cttcgtgatc 52260atggccagtg agaggtcgct cgccatcttc gggacatcca cgacctggcc cggccgccag 52320gaatcggcgg tctcctcggc ggcggcggac atgctgatga cgtagtggtc gagcttgccc 52380cggtggaatc cgggttgcat catccgccgc tggcggcggt gcgagtcccc ggagacggcc 52440acgaggatgg ggccgatgaa ccggctggcg cccgccgcgc ccttgctgcg ggtgaagtcc 52500gccgcgccgg acaccagcat ggtccgcacg atttcggggt gggtggcgag gtagacggtg 52560ttgtggccga ggcggatgcg gaagaggtcc ccgcgttccg tgacggcgga caggaagccc 52620agggggtcgc ggaggagggc cggcaggtgg ccgaggaccg gccaggcacc gggggcctcg 52680gggatggtcg acggaggtga ggacactgtt gctcctgagg ggagggccgg gcgagtcggc 52740gtggggtggg gtgaggtgtg cggtcgggca ggtggtcgcg tcgccggtgg tcggcgacgg 52800gtggtgggtc agggggatcc ggtttcctgg tcgatgagcg cgaacatctc ctcgtccgtc 52860gcctccccga ggtcgggacg cggcgcctcc tccccgccga gcacctgggc gagtgagcga 52920agccgcgacg ccagccggga ccgggcttcc tctcccaggc cctgcgcccc ggggagcggc 52980gacgcgacgg aggagagcac cgcttccagc cggccgatct ccgcgaagag ggactgctcg 53040ggcggcgccg tggccgcgtc gtcggggagg agccgggtca gcaggtgccg ggtgagcgcg 53100gtggcggtgg ggtggtcgaa ggcgagggtg gcgggcaggc gcaggccggt ggcgcgggag 53160agccggttgc ggagttcgac ggcggtgaga gagtcgaagc cgaggtcccg gaaggccgag 53220tccgcaggga ccgcctccgg cgtctgatgg ccgaggaccg tggcgatctg ggtgcgcacc 53280acggtgaaca gggtgtcgtg ttgttgctcg ggggtcaggg tggcgaggcg gtcggcgagg 53340gagacgtcct ggcctgcgcc tgcactggtg ccggtgtgtg cggtgcgggg gctggtgcgg 53400
gcgggcgcga ggtgttccag aaggggcggt gcggggtggg tggggcgtag gtcggcgggc 53460aggagtgcgg gacgtccggt gaccagggcg gtgtcgagga gggcgagggc gtcgggggtg 53520gtcagggggt gcagccccga gcgggtgatg cggtgacggt caccggcgtc cagatgcccg 53580gtcatcccgc tggcctcttc ccacagtccc caggccaggg agagggcggg caggccggcg 53640gcgcggcgct ggtgggccag ggcgtccagg gcggcgttgg cggcggcgta gttgccctgc 53700cccggcgagc ccaggacacc ggccgcggag gagaacagca cgaacgccga caggtccatc 53760cccgcggtca gctcgtgcag atgcagggca ccgtccacct tcgccccgac caccgcatcg 53820atcttctccc ggttcagaca ggccacggtg gcgtcgtcca ggacaccggc cgtgtgcacc 53880acagccgtca gcggatgctc cgcgggcacc tgctccagca gggcggcgac ctgggcgcgg 53940tcggcgacat cgcacgccgc caccgacacc gacacccctg cctgacccag ttccgcacac 54000agttcttcgg caccggtggc ggccatgccg cgccggctca ccagcagcag atgccgcacc 54060ccgtgcccgg cggccagatg gcgcgcgacc gccgctccca gagtgccggt cccacccgtc 54120accagcaccg tcccctccgc atcgaaccgg acggcatccg acgagtcggc gggcaccggc 54180acatgtgcca agcgcgccgc cagcagtcgc ccgccacgca ccgccaactg gggttcgtcg 54240caggccagag ccgccgtgac ggtggtgtcg tcggagaggt cggcatccag caggacgaac 54300cgccccgggt gttccgactg cgccgaacgc agcaggcccc agaccgccgc tccggcgacg 54360tccgtcacct cctcaccggt ccgggtggcc acggcaccgt gtgtcaccac cacgagccgg 54420gcctcggcaa ggcgatcgtc ggccagccag tcctgcacca cgctcagcac ttcgccgagg 54480acgtccgcca ccgcgccctg ggagcacgtc agcagcacgg cgtcggggac gggggcgtcg 54540tcggtgtcca gcccggacag caggccggag aggtccgccg ccgcccggtc atgggtgagt 54600acggtcgacc ggacggccgt gtccgggggc ggggtgcccg gtgtgacgtc cttccaggcc 54660acgtcgaaca gcgccgcgcg gccggccgcc tgggcagagg ctcgcagttc gccggtgtcc 54720
agcggtcgta cggcgagaga gtcgaccgac aacacgcccc ggccggtttc gtcggccagc 54780gacacggaga cggcggtccg ttcgccgtcc cgcccggccg gcgccacccg gacccgtacc 54840gctgccgcct tcaccgcgtg cagggtcaca ccactgaacg agaacggcac cgcccccggc 54900ggcaggcccg tcgccgctcc gagcgccacc gcgtgcaggg cggcgtccag cagagctggg 54960tgcaggttgt accgggacgc ctcgtcgagc acggactccg gcaggcggac ctccgcgaag 55020acctcttcgc cccgccgcca agccgcacgc agcccccgga acgccggtcc gtaggcgaac 55080ccgcgggcct cctgtgccgc gtagaagctt tccagttcgt ccgccgcgca cggcagggcc 55140ccttccggcg gccagctgcg cagggcgtcg ccgtcggcgg agggctgggc gtccagcagg 55200ccggtggcgt ggtgctgcca gggatcctcc gggcgggcgt gctcgctccg ggaggagacg 55260gtgagggtgc gggccccggt gtcgtcgggc gccgacacgc ggacctggag gtcgacggcc 55320gcgtcgtgcg ggacggcgag gggcgcgtga agggtgagct ctcgcacgtg cgcggcaccg 55380ccggcttgga gggcgagttc gaggagggcg gtgccgggga ggaggacgat gccgccgacg 55440ctgtggtcgg cgagccaggg gtgggtgtgc agggacaggc ggccggtgag caaggttccg 55500tctccgtcgg cgagttccac ggtggcgccg aggagagggt gctcggtggg gtgcagtccg 55560gcggcggaga cgtcgccggt gctggtggtg ggtgtgtgga gccagtagtg gtggtgttgg 55620aaggggtagg tgggcaggtc gatggcgggg gtgtgggggg tggtggggta gtgggtggtc 55680caggtgacgg tgtggccggt ggtgtgggcg tgggcgaggg cggtgaggaa gcggtgggtg 55740tcgttgtcgt ggcggcgcag ggtgcccagg gcggtggtgt ggtcgtggtc ttcgatggcg 55800ggggtcaggg tggggtgggg gctggtctcg atgaagaggc ggtggccctg ggcggtgagg 55860gtgcggacgg cgtgggtgaa gcggacgggc cggcgcaggt tgcggtacca gtaggcggcg 55920tccagggtgg tggtgtcggt ccaggcgtct tcgacggtgg agtagaacgg gatgctgccg 55980ggctgggggg tgatgccgtg gagcatgtgg agcaggtcgt gttcgatggt ttcggtgtgg 56040
gggcagtggg atgcgtagtc cacggggatg cggcgggccc gtaccccggt ttcggtgcag 56100tgggtgagga gttcgtccag ggcgtcggtg tcgccggaga cggtggtggc ccgtggcccg 56160ttgaacgcgg ccctccacag ccgtcccggc cagtgggtgg tgatgaggtc ctcggcctcg 56220gttccggtca gggcgagtga ggccatgccg ccgtggcccc gcagcgcggc cagggcccgg 56280ctgcgcagtg ccacgatctt cgcggcgtcc tccagggtga gtgcgccgca gacgtgggcg 56340gcggcgatct cgccctggga gtggccgacc accgcgtcgg gttcgatgcc gtaggaccgc 56400cacagcgccg ccagggacac catgaccgag aacagcaccg gctgcaccac atccgcccgc 56460tcccacaccg gatcctccgc gtcccggtgg agcatttcgg tgagcgacca ctccacccac 56520ggcgccagcg cgcgttcaca cgcaccgaca tggtcggcga acacctggga gtcgcgcagc 56580aggtcgagtc ccatgccggc ccactgaccg ccctggccgg ggaagacgaa caccaccccg 56640ccaccggagt ggctgttggt ctggctgccg gtcgtgtctt cgatgagtgc ggggtgcggt 56700tcacccgagg ccagtgcttc cagcgcgtct tcgaactctg tccgttcccg tgccacgatc 56760acggcgcggt gttccaggac ggccctgcca tgggcgagtt cggctccgac cgcccgtagt 56820gcggtgtcgg tgccggggcc ttggaggaag tcacgcagga gtgcggcctg tgcccgcagc 56880gcggcgtttg aacgcgccga catcgcccag ggcaccgggg cgccgtcttc tggggcgggg 56940tccccggccg gctgggcggg tgcctcttcc aggatgacgt gggcgttggt tccgctgact 57000ccgaaggagg agatgccggc gcgtcggggc cggtccccct ggcgtggcca ggggcggggc 57060tcggtcagca ggctgaccgc gccggacgac cagtcgacct ggggggtcgg ctcgtcgatg 57120tgcagcgtgc gcggcagcgt ctcgtggcgc agtgccatca ccatcttgat gatcccgccg 57180acacctgccg ctgcttgggc gtggccgatg ttggacttca gggagccgag ccagagcggc 57240tggtcggggt ggtgttcctg gccgtaggtg gcgagcaggg cctgggcttc gatggggtcg 57300ccgagtttgg ttcctgtgcc gtgggcttcg acggcgtcga catcggcgcc ggtcaggccg 57360
gagttggcca gggcctggcg gatcacgcgc tgctgggagg ggccgttcgg tgcggtcagg 57420ccgttggagg cgccgtcctg gttgaccgcg ctgccccgca ccaccgccag aacccggtgc 57480ccgttgcgct cggcgtccga caaccgctcc acgagcagca tgcccacgcc ctcgccccag 57540ccggtaccgt cggccgcggc cgcgaaggac ttgcaccgcc cgtccaccga caaaccccgc 57600tgccgggaga agtcgatgaa ggtgcccggt gaagacatca ccgtcacgcc gccggcgagg 57660gccatcgagc attcgcccga tcgcagggct tggcctgcga ggtgcagtgc gacgagcgac 57720gacgagcacg cggtgtcgac cgagacggcc ggcccctcga aaccgaaggt gtaagccacc 57780cgacccgaca cgacactgcc cgcgtttccg ttgccgatgt agccctccgc cccttcggga 57840acggcggtca aacgggcggc gtagtcgtgg tacatcacac ccgcgaacac gcccgttcgg 57900gaaccgcgta cggcagcggg atcgatcccc gcgtgttcga gggtctccca gacggtttcg 57960aggaggagcc gctgctgggg gtccatggca agggcctcac gcgggctgat accgaagaac 58020tcggcgtcga actgcccggc gtcgtagagg aaaccaccgt gccgggtgta cgacgctccg 58080gcccgctccg ggtccgggtc gaacagcccg gccaggtccc acccgcggtc ggccgggaac 58140tccccgatcg cgtcaccgcc cgaagccacc agcccccaca actcctccgg cgaccgcaca 58200ccgcccggga agcggcacgc catcccgacg atcgccagcg gctcgtcact gccgacggct 58260gtggtttcgg cgtacggcga agtgctgtcc gcggcgtcgt ccccgagcag ttccgtgcgc 58320agcaggcggg ccacggccgc ggggctgggc tggtcgaaga ccaggctcgc cggcagtcgc 58380agtcccgtct ccgcgctcag gcggtttcgc agatccacgg ccgtcaggga gtcgaagccg 58440aggtcgcgga aggccgagtc gaccgggatg gcttccggtg cttggtggcc gaggacggtg 58500gcgacatgcg agcggaccag cccgagcagg gcctggtact gctgttcggg tgtccgtccc 58560gcaagccgtg cccgcagcga cgcaccgctg tcagtggtgg ggagggtggt gcggtggctg 58620gtgcgggcgg gcgcgaggtg ttccagaagg ggcggtgcgg gatgggtggg gcgtaggtcg 58680
gcgggcagga gtgcgggacg tccggcggcc agggcggtgt cgaggagggc gagggcgtcg 58740ggggtggtca ggggatgcag ccccgagcgg gtgatgcggt gacggtcacc ggcatccaga 58800tgcccggtca tcccgctggt ctcttcccac agtccccagg ccagggagag ggcgggcaga 58860ccggcggcac ggcgctggtg ggccagggcg tccagggcgg cgttggcggc ggcgtagttc 58920ccctgccccg gcgagcccag gacacctgcg gcggaggaga acagcacgaa cgccgacagg 58980tccatccccg cggtcagctc gtgcagatgc agggcaccgt ccaccttcgc cccgaccacc 59040gcatcgatct tctcccggtc cagacacgtc acggtggcgt cgtccaggac accggccgta 59100tgcaccacag ccgtcagcgg atgctccgcg ggcacctgct ccagcagggc ggcgacctgg 59160gcacggtcgg cgacatcgca cgccgccacc gacaccgaca cccccgcccc acccagttcc 59220gcacacagtt cttcggcacc ggtagcggcc atgccgcgcc ggctcaccag cagcagatgc 59280cgcaccccgt gcccggcggc cagatggcgc gcgaccaccg ctcccagagt gccggtccca 59340cccgtcacca gcaccgtccc ctccgcatcg aaccggacgg catccgacga ctccgacagc 59400ggcggcaccc gcttcaaccg tggtacgcga accaccccgt cgcgtacggc gagttggctc 59460tcaccgcggg ccagagcgga cgcgacggcg gcatcgtcga gaccggcgcc ggtgccgacc 59520ttcgcgtcgg cggacacgtc ggtgtccccg tcggtgtcgg tgtcggtgtc ggtgtcgggg 59580tcggtcttgg tgtcggggtc gaggtccagc aggacgaacc ggtcgggatg ctcggactgg 59640gccgagcgga ccagccccca cacagcggcc cccgccacat cccgcaccgg ctcacccgca 59700tccaccgcga ccgcaccacg cgtcaccacg accagccgcg catccccctc ccgctcatcg 59760gcgagccact cccgcaccac acccaacgcc gcggccgtga cctcggccac cccaccaccc 59820gggcactccc acgccaccaa ctccccgccc ccaccggcca ccgaggccgg caccccctcc 59880accggcaccc accccagctc gaacaacgac ccacgccgca cccgagcccc cagcccctcc 59940aacggcaccg gacgcatcag gagcgactcg agggtgagaa cgggtgctcc ggtctcgtcg 60000
gtggcgtgca ggcgaacggc ggtcgaggtc gcgtcggtgg gagtcatgcg caggcgcagg 60060acggtggcgc cccgggcgtg gagcgagaca ccgttccatg tgtgagggac gagcccggcc 60120tgctgctggt ccgccagcag cagggtgacc gattgcacgg ccgcgtccag gagcgccgga 60180tgcaccccga agcccgcgac atcggccagc ccctcgtccg gcagacgcac ctcggccacc 60240acgtcgtccc cgtcccgcca ggccgcacac aaaccctgga acaccggacc gtagacaaaa 60300cccccacccg ccagacggtc gtagagaccg tcgagaacga ccggtcgggc gccgggtggc 60360ggccactccc cggccgccgc tgcctccgcg ttcggatcgt cttcggtgga cggggacagc 60420acaccctcgg catgccgtgt ccactctccg tccgtctcct cgtccccggc cggccgcgcg 60480tacacattca cggcgcgccg ccccgcctcg tccggcaccg acaccgacac ctgcaccacc 60540acgtgccccg actcgggaat caccagaggg gcgtggagag tgagctcgtc gacacgagga 60600cagccggtac gcagaccggc ctgaaaagcc agatccagga gggcggtgcc cgggaggagg 60660acgattccgc cgacactgtg gtcggcgagc caggtgtggg tgcgcaggga caggctgccg 60720gtgaggacga tcccgtcccc gtccgcgagc tccatcaccg cacccagcag cggatggtcc 60780ggccgctgga gcccggcagc cgacacatcg cccgcaccgg caccgggagt cgcctggagc 60840cagtagtgcc ggcgttggaa ggggtaggtg gggaggtcgg ggatggtgtg ggggtggggg 60900ttggtgtggg tgtggtggtg ggtgtgccag gtggtggggg tgtgggcgag tgcggtgagg 60960aggtggtggg tggggttgtg gtgggggtgg gtgagggtga tggcggtggg ttggtggtgg 61020gggaggttgt ggtgggtgag ggtggtgagg gtgtggtcgg gtccgagttc gatgtaggtg 61080gtgacgccgt tggtgtggag ggtgtggatg gtgtcggtcc agtggacggg ttggcgggtc 61140tggtgggtcc agtagtgggg ggtgaggggg tcgccggggg tgctggtgat gaggggggtg 61200tggggtgggt ggtaggtgag ggtgtgggtg gtggtgtcga gttgttcgag gatggtgtcg 61260gtgtgggggg agtggaaggc gtggttggtg gtgaggggtt tggtggtgat gccttgggtg 61320
tggcaggtgg tggtgatgtg gtggatggtg tcggggtcgc cgctgatgac gagggagtgg 61380gggctgttga cggcggcgat ggtggctttg ccggggtgtt ggtcgaggag gggttggatg 61440tgttcggggg tggtgtggag ggtggtcatg gtgccggggg gcatggtttg catgaggcgg 61500gcgcgggtgg tgatgaggtg ggtggcgtcg gggagggtga ggatgccggc gaggtgggcg 61560gcggtgattt cgccgaggga gtgtccggcg aggtagtggg gggtgatgcc gtaggtttcg 61620gtgatgaggt ggtggagggc tttttggagg gtgaagaggg cgggttgggc gtagagggtg 61680tgggtgagga ggtcggtggg gtcgtggttg aggaggaggt cgcgcagggg gtggtcgagg 61740tgggggtcga agtgggcgca ggtttcgtcg agggcgtcgg cgaaggcggg gtaggtgtgg 61800tagaggccgg tggccatgcc ggggcgttgg gtgccttgtc cggagcagag gaaggcgatt 61860ttgtgggtgt ggccgggttc ggtggggggt gtggggatga ggtgggggtg ggtgcggccg 61920tcggccaggg cggtgagggc gtccaggagg gtgtcgcggt cgggggcgat gagggtggcg 61980cggtggtcga acagggcgcg ggtggtggcc aggctgtagc cgatgtcggc cgcgtcctgt 62040tcggggtggt ccaggacgtg ggcgtgcagg gcggcggcct gggcgcgtag cgcggcctgc 62100gacttgcccg acacgaccca caccaacggc acatccgccg acacccggcc cgcaccgtcc 62160gtctccacca ccaccgccgc agccgccgca gtctccaccc ccgccggcgc ggcttccacc 62220gcctccacct ccgcccgcgc gggcgcctcc tccaggatca cgtgcgcgtt cgtcccgctc 62280accccgaagg acgagattcc cgctcgccgg ggccggtcct cccggcgtgg ccagggccgc 62340gcctcggtca gcaggctcac cgctcccgac gaccagtcca cctgcggcga cggctcatcc 62400acatgcaacg tccgcggcaa cgactcgtgc cgcaacgcca tcaccatctt gatgatcccg 62460ccgacacctg ctgccgcttg ggcgtggccg atgttggact tcagggagcc gagccagagc 62520ggctggtcgg ggtggtgttc ctggccgtag gtggcgagca gggcctgggc ttcgatcggg 62580tcgcccagct tggtccccgt gccatgggct tcgacggcgt cgacatcaac tgcggagagg 62640
ttcgcgttgg ccagggcctg gcggatcaca cgctgctggg acggaccgtt cggcgccgtc 62700agcccgttcg aggcgccgtc ctggttgacc gcgctgcccc gcaccaccgc cagaacccgg 62760tgcccgttgc gctcggcgtc cgacaaccgc tccagcagga gcatcccggc tccctccgac 62820cagccggtac cgtcggccga ggcggagaac gccttgcacc ggccgtccgc cgccaggccc 62880cgctgccgcg agaactccag gaacgcggta ggcgtggaca tcaccgtcgc accgcccgcc 62940aaggccatgg tgcactcgcc cgaccgcagt gcctgacagg ccagatgcag cgcgacgagc 63000gacgacgagc acgcggtgtc cacggacacg gcggggcctt cgagcccgaa cgtgtaggcg 63060acccggcccg acgccacgct tccggacgtg ccggtgagaa cgtacccgtc gacgtcggcg 63120gcgacatggt gccgtgagcg ggacgcgtac tcctgaggca tgacgccggc gaacacgccc 63180gtctggctgc cgcgcacggc accggggtcg atacccgccc gctcgaacgc ctcccacgtc 63240gtctccagca acagccgctg ctgggggtcc atcgcgagcg cctcgcgcgg ggagatcccg 63300aagaatcccg cgtcgaactc ccccgcgtcg tagaggaatc ccccgtgacg ggtgtacgag 63360gtgccccgct gcccgggctc cgggtcgtag agcgcctcca cgtcccagcc acggtcggcc 63420gggaactccc ccaccgcgtc gccgccggag gcgacgagtt gccagaggtc ctcggccgag 63480gcgacacctc ccggataccg gcatcccaca ccgatgatcg cgatgggctc gtgctgcccg 63540gccttcggtt cggcggcggc aggtgccgaa ggcgtcttgg tgtcgttggg gttgaggagg 63600gtggtgaggt ggtgggtgag tgcggtgggg gtggggtggt cgaaggcgag ggtggtgggc 63660aggcgcaggc cggtggcgcg ggtgagccgg ttgcggagtt cgacggcggt gagggagtcg 63720aagccgaggt cgcggaaggt gcgttcgggg tcgatggtgt cgggggtggg gtggcccagg 63780acggcggcga tgtgggtacg ggccagggcc agcagggtgg cgtgccgctg ttcggaggtc 63840agggtggcga ggcggtcggc gagggagacg tcctggcctg cgcctgcact ggtgccggtg 63900tgtgcggtgc gggggctggt gcgggcgggc gcgaggtgtt ccaggagggg tggtgcgggg 63960
tgggtggggc gtaggtcggc gggcaggagt gcgggacgtc cggtggccag ggcggtgtcg 64020aggagggcga gggcgtcggg ggtggtcagg ggatgcagtc ccgagcgggt gatgcggtga 64080cggtcaccgg cgtccaggtg gccggtcatc ccgctggcct cttcccacag tccccaggcc 64140agggagaggg cgggcagacc ggcggcgcgg cgctggtggg ccagggcgtc cagggcggcg 64200ttggcggcgg cgtagttgcc ctgccccggc gagcccagga cacctgcggc ggaggagaac 64260agcacgaacg ccgacaggtc catccccgcg gtcagctcgt gcagatgcag ggcaccgtcc 64320accttcgccc cgaccaccgc atcgatcttc tcccggtcca gacacgtcac ggtggcgtcg 64380tccaggacac cggccgtatg caccacagcc gtcagcggat gctccgcggg cacctgctcc 64440agcagagcgg cgacctgggc gcggtcggcg acatcgcacg ccgccaccga caccgacacc 64500cctgcctgac ccagttccgc acacagttct tcggcaccgg cggcggccat gccgcgccgg 64560ctcaccagca gcagatgccg caccccgtgc ccggcggcca gatggcgcgc gaccgccgcc 64620cccagagtgc cggtcccacc cgtcaccagc accgtcccct ccgcatccag gggcacaggc 64680agggtcagca cgttcttgcc gacatgcagg cccgaccgca tcgaccgcag cgcctggcgg 64740gcctggcgca cgtcccacgc ggtgaccggc aacggctcca gcaccccgcg ccggaacaga 64800tccaccaccg tgtgcaggat ctcccccacc cgctgcgcac ccgcgtccat caggtcatac 64860gcccggtagg acacccccgg gaaccgagcg gcgacctcac cggcatcacg gatgtcggtc 64920ttgcccagct ccaggaaccg gcccccctgc ggcgaacaca gccgcaacga ggcatcggtg 64980tactcacccg ccagacagtt cagcaccaca tccacacccc gcccgccact ggcccggcgg 65040aaacgcgact cgaactccac actccgcgag gaagcgatcc gctgcggcgc gacacccgcc 65100gcccgcagac gcgcccactt cgcctcactc gccgtcgcat acacctccgc ccccagatga 65160cgggcgagct gcaccgccgc cgtaccgacc ccgccggccg ccgcatggac cagcacactc 65220tccccccgcc gcacccccgc cagatcgacc agccccaggt aagcggtagc gaacaccacc 65280
ggcaccgaag ccgcctgcgc gaacgaccag ccctccggga tacgggccag caacacctcc 65340tgcgccacca ccaccggcgc gaacgcgtcc ccgaacaccc cgaacacccg gtctcccacc 65400accaggccct ccaccccggg ccccacctcc accaccaccc ccgcaccctc actgcccacc 65460cccacctgac ccggcaccat ccccaacgcc accagaacat cacggaagtt caccccggca 65520gcccgcaccg ccacccgcac ctgcccccga cccagcacca ccccagccgc atccgaagca 65580accacaccca ccccctccaa caaccccgac ccaccaccat ccagccgcca ccccacccca 65640ccaggcaacg acaacccttc acccgcaccc ccaagccgcc cccaccgctc caaccgctcc 65700aaccgcggca cccgcacgac cccaccacgc acggcaacct gtgcctcgcc acacgcgaca 65760aacccggcca catcgacacc agcaccaaca ccggcgccca tgtcctcatc ggcatcgacg 65820acggtctcca cgccggtgcc ggggtcgagg tccagcagga cgaaccggtc gggatgctcg 65880gactgggccg agcggaccag cccccacaca gcggcccccg ccacgtcccg caccggctcc 65940cccgcatcca ccgcgaccgc accacgcgtc accacgacca gccgcgcgtc cccctcccgc 66000tcatcggcga gccactcccg caccacaccc aacgccgcgg ccgtgacctc ggccacccca 66060ccacccgggc actcccacac caccaactcc ccgcccccac cggccaacga ggccggcacc 66120ccctccaccg gcacccaccc caactcgaac aacgacccac gccgcacccc cgcccccagc 66180ccttccaacg gcaccggacg cagaaccaga gactccagcg cgagcaccag cgcaccggtc 66240tcatcggcaa cccgaagact cacggtcgtt ccggccgcgt cgacagacgt cacccggact 66300cgcagtgccc tggcaccccg ggcgtggagg gaagcaccgt tccatgtgta aggcagcaga 66360ccggcctctt ggtcctcggg cagcaggagg gtgaccgtct gcacggccgc gtccaggagc 66420gccggatgca ccccgaagcc cgcgacatcg gccagcccct cgtccggcag acgcacctcg 66480gccaccacgt cgtccccgtc ccgccaggcc gcacacaaac cctggaacac cggaccgtag 66540acaaaacccc cacccgccag acgaccgtag aactcatcga gatccaccgg ctgcgcaccg 66600
gacggcggcc acaccccgtc cgccaccggc tcaacaacca ccgactcccc aggaacagac 66660ggacacacca caccctcggc atgccgcgtc cactcaccct ccagccctcc gtcctccacc 66720agccgcccgt acacactcac accacgacga cccgcctcgt ccggcaccga aaccgacacc 66780tgcaccacca catgccccga ctccggaacc accagaggag catggagagt cagctcatcg 66840acaccaggac aacccgcacg cagaccagcc tgaaaagcca gctccagcag agcggtaccg 66900ggcagcagga cgacgccgcc gacgctgtgg tcggcgagcc aggggtgggt gtgcagggac 66960aggcgcccgg tgaggacgat tccgtccccg tccgcgagct ccatcaccgc gccgagcagt 67020gggtggtccg gtcgctggag tccggcggcg gagacgtcgc cggtgctggt ggtgggtgtg 67080tggagccagt agtggtggtg ttggaagggg taggtgggca ggtcgatggc gggggtgtgg 67140ggggtggtgg ggtagtgggt ggtccaggtg acggtgtggc cggtggtgtg ggcgtgggcg 67200agggccgtga ggaagcggtg ggtgtcgttg tcgtggcggc gcagggtgcc cagggcggtg 67260gtgtggtcgt ggtcttcgat ggcgggggtc agggtggggt gggggctggt ctcgatgaag 67320aggcggtggc cctgggcggt gagggtgcgg acggcgtggg tgaagcggac gggccggcgc 67380aggttgcggt accagtaggc ggcgtccagg gtggtggtgt cggtccaggc gtcctcgacg 67440gtggagtaga acgggatgct gccgggctgg ggggtgatgc cgtggagcat gtggagcagg 67500tcgtgttcga tggtttcggt gtgggggcag tgggaggcgt agtccacggg gatgcggcgg 67560gcccgtaccc cggtttcggt gcagtgggtg aggagttcgt ccagggcgtc ggtgtcgccg 67620gagacggtgg tggcccgtgg cccgttgaac gcggccgtcc acagccgtcc cggccagtgg 67680gtggtgatga ggtcctcggc ctcggttccg gtcagggcga gtgaggccat gccgccgtgg 67740ccccgcagcg cggccagggc ccggctgcgc agtgccacga ccttcgcggc gtcctccagg 67800gtgagggcgc cgcagacgtg ggcggcggcg atctcgccct gggagtggcc gaccaccgcg 67860tcgggttcga tgccgtagga ccgccacagc gccgccaggg agaccatgac cgagaacagc 67920
accggctgga ccacatcggc ccgctcccac accggatcct ccgcctcgcg gtggagcatc 67980tcggtgagcg accactccac ccacggcgcc agcgcgcgtt cacacgcacc gatatggtcg 68040gcgaacaccc ccgaggtcgt cagcagatca agtcccatgc cggcccactg accaccctgg 68100cccgggaaca cgaacaccac cccgccaccg gaatggctgt ggctgccggt cgcgtcttcg 68160atgagtgcgg ggtgcggctc acccgaggcc agtgcttcca gcgcgccttc gaactccgcc 68220cgctcccgtg ccacgatcac cgcgcggtgc tccagcacgg ccctgccacg agccaactct 68280gccccgatat cccgcacccc ggcatccgta ccggggccgc gcaggaactc acgcaagacc 68340atggcctgcg cccgcaacgc cgcacccgaa cgcgccgaca ccacccaggg caccggagcc 68400ccgtcctcta ccgcagcctc cccgggccga cgggcgggtg cctcctccag gatcacgtgc 68460gcgttggttc cgctcacccc gaacgaggac accccggccc gccggggccg gtcctcccga 68520cgcggccagg gccgcgcctc ggacagcagg ctcaccgccc ccgacgacca gtccacctgc 68580ggtgacggct catccacatg caacgtccgc ggcagcgact cgtgccgcag cgccatcacc 68640atcttgatga tcccgcccac acccgctgcc gcctgggcgt ggccgatgtt ggacttcacc 68700gagcccagcc acaacggctg ttccccggaa cgctcctggc catatgtgtc gagcagggcc 68760tgggcttcga tcgggtcacc gagccgtgtg ccggtcccgt gcccctccac cgcgtcgacg 68820tccgccaccg tcagccccgc gttggccagt gcctcgcgga tcacgcgctc ctgcgagggg 68880ccgttcggtg cggtcaggcc gttggaggcg ccgtcctggt tgaccgcgct gccccgcacc 68940accgccagaa cccggtgccc gttgcgctcg gcgtccgaca accgctcgac cagcagcatg 69000cccacgccct cgcccatgcc ggtaccgtcg gccgcggccg cgaaggactt gcaccgcccg 69060tccaccgaca gaccccgctg ccgggagaac tccacgaaca ggagcggggt ggacatcacg 69120gtgaccccac cggcgagggc gagatcgcac tcgcccgtcc gcagcgactg gcaggccagg 69180tgcagcgcca ccagcgacga cgaacacgcc gtgtcgacgg tgacggccgg gccttccaga 69240
ccgagcgtgt aggcgacgcg cccggaggcg acggcgccac cgctgccgtt gccgatgtag 69300ccctcgaacc cttcggggat ggtggcgagc cgggaggcgt agtcgtggta catcatgccg 69360gtgaagacac ccgctcgggc tccccggacc gaggaggggt cgatcccggc ccgctcgaag 69420acctcccacg aggtctccag cagcaagcgc tgctgggggt ccatggccag ggcctcacgc 69480ggactgatgc cgaaaagctc ggcgtcgaac tgtccggcgt cgtagaggaa accaccgtgg 69540cgggtgtacg acgctccggc ccgctccggg tccgggtcga acagcccggc caggtcccac 69600ccgcggtcgg ccgggaactc cccgatcgcg tcaccgcccg aagccaccag cccccacaac 69660tcctccggcg accgcacacc gcccgggaac cggcacgcca tcccgacgat cgccagcggc 69720tcctgtgccg cctcgacggc cgctgtgagc tgctggttgc gccgccgcag ggcctcattg 69780gccttcaggg atgcccgcag cgcctcgacg agcttctcgc tgggcgtagc catcggtgtc 69840tccaagtctg cgaatccggc aggtgcggac gcggtggtgt ggacggggcg ggggtcggcg 69900gggaccgcgg cgggcgactc gggtggtgtc agcgacgccg ctgctcggtg agcccggcca 69960gccaggtgtg gacgtgccgg gccgtcgact ccgcgtgctc ttcgagcatc gtgaagtggt 70020tgccgtcggt ttcgaggacg gtgtgcggct cgccccacac cggcggcggc tgttcgctct 70080cacgggcgcg gaggaagagg gtgggtgtct cgagggcggg cggccgccag cccgcgaaga 70140tgcggaagta gccgcccatc gccaccaggc gggcgtagtc caggtcgatg aactcggtga 70200cgcggtcgaa gatttcgctg gtgagggcgg cggcgacggg ggccatcccc tcgtcgggca 70260ggtaggcgtc catgaccacc acggcctgcg gccggacgcc caggtgttcc aggcggctcg 70320tgacggtgtg ggtgaaccag ccgccggcgg agtgtccggc gagggcgaag ggctcgccgt 70380cggtgtggcg gaggatggcg tcggtgaaca gccgggtgat ggtgtcgacg tcggcgggga 70440ggggctcgcc gtcggcgaag ccgggcgccg gcacgtacca gacgtcgcgg agcccgtcga 70500gggccgccgc gaagcgggag tactggtaga cgctggacac ggcggcgacg gtgggcaggc 70560
agatcagcgc gggcccggtg tcgccctggg cgacgcggac gaaggggggt cgggtcatag 70620ccgaggggtc ggtgaagcag ggccggaagg cggaggccgc cgacagcagg gccatggact 70680cctcgacgcg gccgctgtcg tgaccgatcc agaacagggc ttccaccgtg tcggcggacg 70740ggccgctccc ggctcgcgac gaggcggtgg catcgcgctc cccaggggcg ccggccgttt 70800cggcggtcat atcggaggcc ggctcggcgg cgaggagcct ttcgaggtgg tcggcgagcg 70860ctgccggggt cgggtggtcg aagacgagcg tggtggccag gcgcagcccg gtcgctgcgt 70920tgaggcggtt gcgcagttcc acggcggtca gggagtcgaa gccgaactcg cggaactcgc 70980cgtcggcggt gacggtgtcg gtgccgccgt ggccgaggac ggccgcggcg tgggtgcgga 71040ccacctccgt cagcagggcg gtgcgctcgg cgggcttcgg ggtcccggcc agtcgcccgc 71100ggagttcggc ggcggcgtcg gccccgacgc cgtggtcggc ggtccggcgg gccggggtgc 71160ggaccaggcc cctgaggacg ggcggcaggg tgccgacggc cgcctgctca cggagggtgc 71220ccgggtcgag gggggtggcg aggagcagcg gttcgtcgag ggcgagggcg gtgtcgaaca 71280gggcgagccc gtgggcgttg gtgagcggga gcaggccgct gcgggtcatc cgggcgacgt 71340cggcggcggc gaggtgctcg gccatgccgc ccgcctcggc ccagcgtccc caggcgaggg 71400agcggccggg caggccgagg gcgtgccgtt gctgcatcag agcgtcgagg aaggcgttcg 71460cggccgtgta gttggcctgt ccggggctgc cgaaggaggc ggcggcggac gagaaggcga 71520tgaacgcgtc gagcccggcg tcgcgggtga ggtcgtgcag gtgggcggcg ccgtgggcct 71580tggcgctcag gacggcgtcc aggcggtcgg gggtcaggga ggtgaggacg ccgtcgtcga 71640cgacgccggc ggtgtggagc acggccttga gcggatgccg tgccgggatc tcggccagca 71700gcgccgcgac ggcccgccgg tcggcgaggt cgcaggcgac ggccgtcgtc cgggcgccca 71760gctcggcgag ttcggcgacg agttcggcgg tgccgggagc ggtggggccg ctgcggctgg 71820tcagcagcag gtgccgtacg ccgtgggtga cgacgaggtg gcgggcgagg agccggccga 71880
ggtagccggt gccgccggtg atgaggacag tggcgtcggg gtcccagtgt ccgctgtccg 71940ctcgggcgcc gaccgggatg cgggccagcc gcggggtgtg ggcgcggccc tcgcgcagga 72000cggtctgcgg ctcaccggag agcagggccg cggccagggc gcgccggctg gcgtcggtgt 72060cgtcgaggtc ggtgaggacg aaccggccgg ggttctcggt ctgggcggag cggaccatgc 72120cccagacggc ggcgtgcgcg aggtcgggga cggagtcgcc cggtgcggcg gcgaccgcgc 72180cgtgggtgac gaacgcgagc cgggagtccg cgaaccggtc gtcggcgagc cagctctgca 72240gcaggtgcag gacgcggacg gtggcccgcc gggtggcgtc ggccgcgtcg gcggcgccgt 72300cgcggtgcgg gcaggggacg acgaccacgt cgggtacggg tgtgccggcc gaggccagtt 72360cctccagatc cgcgtatgtg ctccacggca cgccgggggc gtcggggcac tcggcttcgg 72420agccgatcag cgccaggcgc gtcttcgacg acggtgtcct gggcagcggt acgggcgccc 72480agtcgagccg gaagagggcg tcgtggtggg cggtgcgggc cgagtggagc tgtccggccg 72540tgacgggccg gaacgcgagt gactcggccg tgacgacggt gtgtcccgtg ctgtccgtgg 72600ccagcagggc gatcgtgtcg ggcgaccgcc gactgaggcg gacgcgcagc gccgatgccc 72660cggaggccgt gacggtgacg ccgctccagg agaagggcag ccagccgtgg ccctcgtccg 72720gctcgtcctc ggcgaagccg aggaccaccg ggtggagtgc cgcgtcgagc agcgccgggt 72780ggacggcgta gcggtcggcg tcgcccgacg gtccgtcggg cagtgcgacc tcggcgtaca 72840ggtcgtcccc gtgccgccag gcggcccgca gtccctggaa cgcgggtccg tatccgaggc 72900ccgcgtcggc cagtgtcccg taccagtggt cgaggtcgac ggggaccgcg tccgtgggcg 72960gccacggtgc ggcggtgtcg tgggcggtct ccgcgcgccg tgtcaggacg cccgtcgcgt 73020ggcaggtcca gccggtcccg tccgtgccgg tcggcgcggc gggggtgagt ccgtcgtctt 73080cccgcgcgta gagcgtgaag gggcgccgct ccaccccgtc cggcgccgtc tcggtcgcgc 73140cgacggagag ctgcaggacg accgatccgc gctccggcag gaccagcggg acctggagtg 73200
ccagttcctc gacggtgtcg cagccgactt cgtcgcccgc gcggacggcc agttcgagga 73260tggccgtgcc gggcagcagg acggtgccga agacggcgtg gtcggcgagc cagggatggg 73320tgcgcagcga gatcctgccg gtgagcaggc attcctgcga ctgcggtgat ccggccaggg 73380gtacggcgga gccgagcagg gggtgtccgg ctgcggtgag tccggcggcc gagacgtccc 73440cggacaggct ggtgtcggcg tccagccagt agcggcggcg gtcgaagggg taggtcggca 73500ggtcgaggtg gcgggcgcgt tccggtgtcg cgccgatgag ggccggccag tggacggccg 73560tcccgccctt cggtgtgccc tgcacgtgga ggtgggcgag cgcggtgagc agcgccaggg 73620gttcggggcg gtcggcgcgc agcagcggga ccagggcggg accgggctcg gtggtgttgt 73680cgtcggcggg caggcactct ccggcgaggg cgcagagggt tccgtccggg ccgagttcca 73740ggaaggtgcg taccccgtcg tcgtcgtgga ggcggcgtac ggcgtcgccg aagcgtacgg 73800tgcggcgtag ctggcggacc cagtactccg ggtcggtgag cgtgccggcc gtggcgcggt 73860cgccggtgac ggtggagacc accgggatcg tcggttcggc gtaggcgatg ccggtcgcga 73920cctgccggaa ctcctccagg atcggttcca tcagcgggga gtggaaggcg cggtcggtgc 73980tgaggcgttt ggtgcgcagg ccctgctcgg cgaaggcggc ggcggcttcc agtacgtccg 74040gctcggctcc ggagatcacc accgacgtgg ggccgttgac agcggcgacg gccacccgtg 74100cctcccggcc ggcgagcatc cgggtgacct gttcctcgct cgcgcggacc gccagcatgg 74160ctccgccggg cggcagttgt gtctgcgcca gccggccccg ggccgcgacc agccgggccg 74220cgtcggtcag tgagaggacg ccggcgacgt gcgcggcggc cagttcgccg atcgagtggc 74280cggcgacgtg gtccgggcgg atcccggcgc tctccagcag gcggaagagg gcgacctgga 74340gggcgaagag cgccggctgc gcgtcgccgg tgcggtccag cggctgcggc tcgtcgagga 74400ggagcgggcg caggggccgg tcgagatggg gttcgagctc cgcgagtacg tcgtccaacg 74460cctgggcgaa ggcggggtgg gcggcgtaca gctctcggcc catgccgggc cgttgggttc 74520
cctggccgga gaagaggaag gcgaccttgc cgtgggcggt gcgggcgggg gattcgatca 74580ggccgggggc cgtggtgccc tcggccagtg cgtccagggt gcgcaggagt ccctcgtggt 74640cctcggcgac cagcacggcc cggcgttcga atgtcgaccg gccggtggcc agggcgtgtg 74700cgacgtcacc gatcgggatg tcggggttgg cggcgaggta gtcgcgcagc cgcctggcct 74760gggcgcgcag ggcggtgtcg gtcctggccg agaggagaca gggcaccgtc gcgggtccgg 74820cctcgtcctg cgacggtgcc tcctccggcc gtacctcttc ctcctgcggt gcctcttcca 74880ggatgacgtg tgcgttggtg ccgctcaccc cgaacgacga cacacccgca cgacgcggac 74940gctcaccccg ctcccacacc acctcctccg tcagcaaccg caccgcaccc gacgaccaat 75000ccacatgcgg cgacggctca tccacatgca acgtccgcgg cagacgaccc cgccacaacg 75060ccatcaccat cttgatcaca ccagccaccc ccgccgcagc ctgcgtatga cccagattcg 75120acttcaccga ccccaaccac aacggcaccc cacgaccccg cccatacgcc gccagcaccg 75180cctgcgcctc gatcggatca cccaacgacg tccccgtccc atgcccctcc accacatcca 75240cctcagcagc cgacaacccc gcacacacca acgcctgacc gatcacccgc tgctgagacg 75300gaccattcgg cgccgtcaac ccattcgacg caccatcctg attcaccgca ctcccccgca 75360ccaccgccaa cacccgatgc cccagccgcc gcgcatccga caaccgctcc accaacaaca 75420cacccacacc ctcggaccac cccaccccgt ccgccgccgc cgcgaacgcc ttgcaccgcc 75480cgtccgccga caaaccccgc tgccgcgaga actccacgaa cgcccccggc gtcgacatca 75540ccgtcacacc ccccgccaac gccaactccg actcacccga ccgcaacgac tgacacgcca 75600gatgcaacgc caccaacgac gacgaacacg ccgtgtccac cgtcaccgcc ggaccctcca 75660acccgaacgt gtacgacaac cgccccgaca acacactccc cgacacaccc gtcagcgcat 75720acccctccag atcccggcca ccccgacgca ccaactccgc ataatcctga ttcgccaccc 75780ccgcgaacac acccgtacga ctcccccgca acgacaccgg atcgatcccc gcccgctcca 75840
gcgcctccca ggacacctcc agcaacaacc gctgctgcgg atccatcgcc aacgcctcac 75900gcggcgaaat cccgaaaaac cccgcatcga actccgccgc acccgccaaa aacccacccg 75960accgcgtata cgacgacccc ggccgccccg cctccggatc gtaaagaccc tccacatccc 76020agccccggtc caccgggaag ccgccgatcg cgtccccacc cgaggcgacc aactcccaca 76080aatcctccgg cgaccacaca ccccccggaa aacgacacgc catccccacg atcgccaccg 76140gctcatccac cacaccgggt cgggccgcga cgggcggtgt cgccggggcg gttccgcaca 76200gctggtcccg gaggtgccgg gccaggacgg cggggcgcgg gtagtcgaag accagtgtcg 76260tgggaaggcg cagtccggtg gccgtgttga ggcggttgcg cagttcgacg gcggtgagcg 76320agtcgaagcc gagttcgcgg aaggcccggt cggccgggac ggtgtcggcc tcccggtggc 76380cgagcaccgc ggccgtgtgg gtgctgacca gttccagcag gacgtccgtc cgggcgtccg 76440gttccagggc ggcgagccga tcgcgcaggt cggtgccgtg ggcggtgccg gtgccggtgg 76500cgcgggtgag cgcccggacg tcggggaggt cgccgatcag gggcaggcgg ctgccggcgg 76560tgtgggcggt gaagcgctcc cagtcgatgt cggcgacggt caagccgctc tcgtcgtggt 76620ccagcacccg ggccagggcc gccagggcga gttcgggcgc catctccgtc agtccccggc 76680ggtgcagccg ggcggccgcg tccggccggc cggcggcgct gtgtccgcgc caggggcccc 76740aggccaccgc ggtggaaggc agtccgagac cgcgccggtg gacggcgaga gcctcgacat 76800aggcgttggc cgccacatag gcaccctggc caccggaacc gaacgtggcg gcggccgagg 76860agaacaccac gaacgccgaa agatccgcac cccgcgtcag ctcgtgcagg ttccgcgcac 76920ccaccgccct ggccgccagc accccctcca gccgctccgg cgtcaacgcg tccagcacac 76980cgtcgtccac cacccccgcc gtatgcacga ccgcccccag cggacaatcc tcgggaaccg 77040ccgtggccag cagctccgcg agcgcccccc gatcggccac atcacaggcg gcgatggtca 77100cccgggcgcc ccactggtcc gtcgtgtcgg cgaggccgaa gccgtcgccg gaggtactga 77160
tcagcagcag gtgttcggct ccgcggtcgg ccagccatcg ggcgaggtgg gcggccggct 77220gctcggggtc ggcgttctcg ccggtgatca ggacggtgcc gcgcggccgc cacccaccag 77280cctccgctcc ccctcccgga gcacgcacca gacgccgcac gaacaccccc gacgcccgga 77340ccgcgacctc cccctcaccc ccgccccccg acagcacacc cagcaaaccc tccaccaccc 77400gctcgtcgac cacctccggc agatcgacca ccccacccca ccgatccggc aactccaacc 77460ccgccacccg gcccaaaccc cacaccacag ccccacccgg atcccccagc cgatccccct 77520cccccaccga caccgcaccc cgcgtcacac accacaacgg cacccccaac ccctccaccg 77580cctggaccaa ccccagcacc aacccggcaa cacccacccc acccccacac acagccagca 77640cccccaccgg cccctcacca ccccacacct cacgcaaccg ctcccccaac accacccgat 77700ccgcacaacc cccctccaca gccaccaccc gcacacacac ccccgcccac tccaaaccct 77760ccaccacagc agcagcaccc accaccccct caggcaccac caccacccac accccacccg 77820acaccacacc accacccgac cgcgacaccg gacgccacac cacccgatac cgccagccat 77880cgaccaccgc acgctcccga acaccccgac gccagtcgcc gagcgcggac accagcgcgt 77940cgagcggcgc gtcctcgtcc acggcgagca gcgcggccac ggccgccggg tcctctcgtt 78000cgacggcttc ccacagcggg ccgtcctccg tggtggccgg cgcggccggt gtctcctccg 78060ggtccagcca gtaccgctca cgctcgaagg cgtacgtcgg cagctccacc cggccggcgg 78120tcccggacgg tttgccgccc agtacggccg cccagtccac ccgtaccccg cgcacggaca 78180gctccgccgc ggaggccagg aagcgccgca gaccgccctc gccccggcgc agtgagccga 78240ccaccagggt gtcggcggcg ccgaggtcgt cgagcgtctc ctggaccgcg acggagaccg 78300cggggtgcgg gccggcctcg acgaagacgg tgtgcccgtc gcgggcgagg gcccgggtgg 78360cgtcccggaa ccggacgggc tcgcgcaggt tgcggtacca gtacgcggcg tcgagtgcgg 78420tgccgtcgac gggctcgccg gtgaccgtgg agtagagcgg gatgtcggcg gggcgggggg 78480
tgacgggagc gagaaggccg agcaggtctg cgcggatcgc ctcgacctgc ggggagtgcg 78540aggcccagtc gaccttcagc aggcgggccg ggacgccgtc ccgggtcagg tcgtcgacca 78600gggcggtgac cgcgtccggg gagccggaga ccacggccga gcgggccccg ttgtcggcgg 78660cgaccaccag gctcgggtcc acggcggcga gccgcggttc caggtcctcg gccggcagac 78720cgaccgaggc catggccccc tgtccggcga gcgcggcgag ggcctggctg cgcagggcgg 78780tgacgcgcgc cgcgtcctcc agggagaggg caccggcgac gcaggccgcc gcgatctcgc 78840cctgggagtg tccggcgacg gcgtcggggc ggacgccgta ggagcgccag agggccgcca 78900gggacaccat gaccgcgaag agcacgggct ggacgacgtc gacccggtcc agcggcgggg 78960cgtccggttc gccgcgcagg acgtcgagca gttcccagtc gaggtacgga cgcagggcgt 79020cggcgcattc ggtcatgcgc tgggcgaaga ccggtgaaga gtccaggagt tcggcggcca 79080tgccgtccca ctgggtgccc tggccgccga agagcagcgc gattttgccg tccgcctcgg 79140ggccggtgcg tccggccacg actccggccg tcggcaggcc ggtggcgagg gcgtcgaggc 79200cgtgccggaa accgtcgagg tcctcggcga gcacgaccgc ccggtgctcc agccacgccc 79260gctccaccgc cagcgcacgc ccgacctcca ccggagccgc ccccgcccca tcggcgaaca 79320cccgcaaccg ccgcgcctgc ccccgcaacg ccgactccga acgagccgac accacccacg 79380gcaccaccgc gggtccggcc tcgccctgcg acggtgcctc ctccggccgt acctcttcct 79440cctgcggtgc ctcctccaga atcacatgcg cgttggtgcc gctcaccccg aacgacgaca 79500cacccgcacg ccgcgggcgc tcaccccgct cccacaccac ctcctccgtc agcaaccgca 79560ccgcacccga cgaccaatcc acatgcggcg acggctcatc cacatgcaac gtccgcggca 79620gacgaccccg ccacaacacc atcaccatct tgatcacacc agccaccccc gccgcagcct 79680gcgtatgacc cagattcgac ttcaccgacc ccaaccacaa cggcacccca cgaccccgcc 79740catacgccgc cagcaccgcc tgcgcctcga tcggatcacc caacgacgtc cccgtcccat 79800
gcccctccac cacatccacc tcagcagccg acaaccccgc acacaccaac gcctgaccga 79860tcacccgctg ctgagacgga ccattcggcg ccgtcaaccc attcgacgca ccatcctgat 79920tcaccgcact cccccgcacc accggcaaca cccgatgccc cagccggcgc gcatccgaca 79980accgctccac caacaacaca cccacaccct cggaccaccc caccccgttc gtcggcgtcg 80040cgaacgcctt gcaccggccg tccggcgaca aaccccgctg tcgcgagaac tccacgaacg 80100cccccggcgt cgacatcacc gtcacacccc ccggcaacgc caacttcgac tcacccgaac 80160gcaacgactg acacgccaga tgcaacgcca ccaacgacga cgaacacgcc gtgttcaccg 80220tcaccggcgg acccttcaac ccgaacgtgt acgacaaccg ccccgacaac acactccccg 80280acacacccgt cagcgcatac ccctccagat cccggccacc ccgacgcacc aactccgcat 80340aatcctgatt cgccaccccc gcgaacacac ccgtacgact cccccgcaac gacaccggat 80400cgatccccgc ccgctccagc gcctcccagg acacctccag caacaaccgc tgctgcggat 80460ccatcgccaa cgcctcacgc ggcgaaatcc cgaaaaaccc cgcatcgaac tccgccgcac 80520ccgccaaaaa cccacccgcc cgcgtatacg acgaccccgg ccgccccgcc tccggatcgt 80580aaagaccctc cacatcccag ccccggtcca ccgggaagcc gccgatcgcg tccccgcccg 80640aggcgaccaa ctcccacaaa tcctccggcg accacacacc ccccggaaaa cggcacgcca 80700tccccacgat cgccaccggc tcatccacca caccggaccg gatgaaggcg ggccggccgg 80760ccggggcttc cccgccggtg ctcagcagtg tgccgaggtg tgtggccagg gcggacgggt 80820tggggtagtc gaacaccagc gtgctgggca accgcagtcc ggtggccgtg ttgaggccgt 80880tgcgcagttc gacggcgttg agcgagccga agccgagctc ccggaaggcc cggtcggccg 80940ggacggcggt ggcggtgcgg tggccgagga cggtggcggt gtgggtgcgg acgaggtcga 81000gcagggcgcg gtcccgttcg gccggttcca gggcggccag gcgtgcgcgg agcgagccgg 81060gggcctccgt gccggtggcg ggccgggcga gccgggcctc ggggatgtcg gagagcagcc 81120
gggcgaggcc gtcggcggcg ggcaggcgct cccagtcgat gtcggcgatg gtgaggcagg 81180tctcgttgcg gtccagtacc tggccgaggg cggacagcgc gggctcggtg tccatgggcc 81240ggatcccgcg gcggtccatc cgcgtggcgg cctccgcgtc cgcggccatg cccccgcccg 81300cccaggcgcc ccaggccacc gcggtggagg gcagtccgag accgcgccgg tggacggcga 81360gagcctcgac ataggcgttg gccgccacat aggcaccctg gccaccggaa ccgaacgtgg 81420cggcggccga ggagaacacc acgaacgccg acagatccgc accccgcgtc agttcgtgca 81480ggttccgcgc acccaccgcc ttggccgcca gcaccccctc cagccgctcc ggcgtcaacg 81540cgtccagcac accgtcgtcc accacccccg ccgtatgcac gacggcaccc agcggacaat 81600cctcgggaac cgccgtggcc agcagctccg cgagcgcccc ccggtcggcc acatcacagg 81660cggcgatggt cacccgggcg cccatcgcgg tgagttccgc gcggagctcc ccggcaccct 81720tggcctcgcg tccgctccgg ctgaccagca gcaggtgctc ggccccgcgc cggaccatcc 81780agcgggcgac gtgtgctccc agagcgccgg tgccgccggt gatcaggacg gtgccgcgcg 81840gccgccaccc accagcctcc gctccccctc ccggagcacg caccagacgc cgcacgaaca 81900cccccgacgc ccggaccgcg acctccccct cacccccgcc ccccgacagc acacccagca 81960aaccctccac cacccgctcg tcgaccacct ccggcagatc gaccacccca ccccaccgat 82020ccggcaactc caaccccgcc acccggccca aaccccacac cacagcccca cccggatccc 82080ccagccgatc cccctccccc accgacaccg caccccgcgt cacacaccac aacggcaccc 82140ccaacccctc caccgcctgg accagcccca gcaccaaccc ggcaacaccc accccacccc 82200cacacacagc cagcaccccc accggcccct caccaccaca cacctcacgc aaccgctccc 82260ccaacaccac ccgatccgca caacccccct ccacagccac cacccgcaca cacacccccg 82320cccgctccaa accctccacc acagcagcag cacccaccac cccctcaggc accaccacca 82380cccacacccc acccgacacc acaccaccac ccgaccgcga caccggacgc cacaccaccc 82440
gataccgcca gccatcgacc accgcacgct cccgaacacc ccgacgccag tcgccgagcg 82500cggacaccac ggatcccaaa ggcgcgtcct cgtcgacttc cagcagggcc gcgaccgccg 82560gcaggtccgc gcgctcgacc gcctgccaca gcgggccgtc ctccccggcc ggcagcgcgg 82620ccggtgtctc gcccgcgtcc agccagtacc gctcacgctc gaacgcgtac gtcggcagct 82680ccacccggcc ggcggtcccg gacggtgcgc cgcccagcac ggccgcccag tccacccgca 82740ccccgcgcac ggacagcccg gccacggcgg cgagcacgga cacggcctcc ggccggtccg 82800gtcgcagtgc ggggagcagg ggggcgggtt cggtgagggc gtcctggccg agggcgcaga 82860gtgtgccgtc cgggccgagt tcgaggtagg cggtgacgcc ctgggcctgg agccaggcga 82920ggccgtcgcc gaagcggacg gtgtggcggg cgtgctggac ccagtagtca gcggtgccca 82980tggtgtcggc ggagacgggg gcgccggtga ggttggtgac cacggggatg cgcggcgggg 83040cgaagacgac ctgctccgcg acgcggcgga agtcgtccag tacggcgtcc aggtgcgggg 83100agtggaaggc gtggctggtg cgcagccgcc gggtccggcg gccctgttcc gcccagtggc 83160gggcgagcgt gagtacggcg tcctcgtcgc cggcgaggac gaccgcgcgc gggccgttga 83220cggcggccag gtccgcgcgc ccctcggcat cctggagcag cggccggact tcctcctccg 83280tcgcctcgac ggcgaccatg gcgccggtgt ccggcagcgc ctgcatcagc cggccccggg 83340ccgtcaccag ggcggccgcg tcggggaggg agagcatccc ggcgacgtgt gcggcggcca 83400gttcaccgac ggagtgcccc aggaggtagt cgggtgtcac cccccagttc tcgaccagcc 83460ggtacagcgc gacctcgacg gcgaacaggg cgggctgggc gtattccgtc tgttcgatca 83520gctcggcgcc gggggatccg ggggccgcga acacgatgtc gcgcagggtg tggcctgctt 83580ccccgatcgg gccgaagtgg gcgcacacct cgtcgaaggc gtccgcgaag gcggggaagt 83640gcgcgtggag ttcgcggccc atggccgggc gctgtgtgcc ctggcccgcg aagaggaacg 83700ccagcgggcc ttcgtcggtc gcggtgccgg tgacgacttc cggggcggga cggccggtgg 83760
cgagggcgtc gaggccgtgc cggaaaccgt cgaggtcctc ggcgagcacg accgcccggt 83820gctccagcca cgcccgctcc accgccagcg cacgcccgac ctccaccgga gccgcccccg 83880ccccatcggc gaacacccgc aaccgccgcg cctgcccccg caacgccgac tccgaacgag 83940ccgacaccac ccacggcacc accgcgggtc cggccccgtc ccccgacgga accaccaccg 84000gcccgacgcc gtcccccgac ggtgcctcct ccggccgtac ctcttcctcc tgcggtgcct 84060cctccagaat cacatgcgcg ttggtaccgc tcaccccgaa cgacgacaca cccgcacgcc 84120gcggacgctc accccgctcc cacaccacct cctccgtcag caaccgcacc gcacccgacg 84180accaatccac atgcggcgac ggctcatcca catgcaacgt ccgcggcaga cgaccccgcc 84240acaacgccat caccatcttg atcacaccag ccacccccgc cgcagcctgc gtatgaccca 84300gattcgactt caccgacccc aaccacaacg gcaccccacg accccgccca tacgccgcca 84360gcaccgcctg cgcctcgatc ggatcaccca acgacgtccc cgtcccatgc ccctccacca 84420catccacctc agcagccgac aaccccgcac acaccaacgc ctgaccgatc acccgctgct 84480gagacggacc attcggcgcc gtcaacccat tcgacgcacc atcctgattc accgcactcc 84540cccgcaccac cgccaacacc cgatgcccca gccgccgcgc atccgacaac cgctccacca 84600gcagcacacc gacaccctcg gacatcccgg tgccgtcggc cgcggcggcg tagggcttgc 84660agcggccgtc cgccgacagg ccccgttggc gcgagaactc cacgaacatg gcgggggtgg 84720acatcacggt gaccccgccg gcgagtgcga gggaggattc gcccgacctg acggactggc 84780aggccaggtg cagtgccacc agcgatgacg agcacgccgt gtcgaccgtc accgcggggc 84840cttcgaaccc gaaggtgtag gagagccggc cggacaggac gctcgccgcg ttgccgttgc 84900cgaggaagcc ctgaaggtga tccggaacgg acagcagacg ggtggcgtag tcgtgcgaca 84960tcatccccgc gaagacgccg gtgcggctgc cgcgcagggt ggccgggtcg atcccggccc 85020gctccagcgc ctcccaggac acctccagca tcaaccgctg ctgcgggtcc atcgccagcg 85080
cctcgcgcgg ggagagaccg aagaatcccg cgtcgaactc cgctgcctcg tgcaggaatc 85140cgcccgatcg cgtgtacgac cgccctgccc ggcccggctc cgggtcgtag aggtcctcga 85200cgtcccagcc ccggtccacc gggaagtcgc cgatcgcgtc cccgcccgag gcgaccagct 85260cccacaggtc ctcgggcgat cgcacacctc ccgggaaccg gcacgccatg cccacgatcg 85320cgaccggctc ctgcctgccc gactcgacct gctccagccg gcgccggacc cgcagcagat 85380cggcggtcgc gcgcttgagg tactcgcgga gcatttcctc gttggccatg acggggtctc 85440ctcgccgctg cgctggaggt ggcacggaac cccgccagat tagggtgggc aagtcaaccc 85500gaataccccc tatacacccc agactggcta cgtgaagcga atacccgttc aaataggggg 85560aagagccgca ggcatggatc gttacgcgaa gcgtttcgag gaccggctgg tcctggtcac 85620gggggcgggg agcggcatcg ggcgggcgac ggcctgccgg ttcggtgccg ccggggcgcg 85680gctggtgtgt gtggaccggg acgggcccgg cgcggaggcg accgccgaac tggcgcgtgc 85740gcggggggcg cgggcggcgt gcgccgaggt ggccgacgtc tcggacgagg tggcgatgga 85800gcggctcgcc gcgcgcgtca cggccgcgca cggcgtgctg gacgtgctcg tgaacaatgc 85860cggtatcggc atgtcggggc ggtttctcga cacgtcggcc gaggactggc gccgcaccct 85920gggggtgaat ctgtggggcg tcatccacgg gtgccggctc ctcggccggg gcatggccga 85980gcgccggcag ggcggtcaca tcgtgacggt ggcctcggcg gccgcgttcc agccgacccg 86040ggtcgttccg gtgtacgcca ccagcaaggc cgcggccctg atgctgagcg agtgtctgcg 86100cgcggagttg gcggagttcg gcatcggtgt gagcgtggtc tgccccggcc tggtccgtac 86160gccgttcgcg tccgcgatgt acttcgccgg cgcgtccccc gacgagcaca cccggctgcg 86220tgagtcctcc gcccgccgct tcgcgggccg cggctgcccg ccggagaagg tcgcggacgc 86280cgtcctgcgc gcgatcatgc ggacggcctt gccgacggtg accgggtcga cgccgtagag 86340ctggatcagc gcggtctcct cgcccgtctc cggcttgacc tcgaagtacg cgagcggctc 86400
ggcgtcggcg gctgccgcgt cgtacagcag gatgcgcaga tccggaagtc ctgctcttcg 86460acgagccgtt cagcgcgctg gacccgctga tccctttagt gagggttaat tgcggccgcg 86520ttccagccga cccgggtcgt tccggtgtac gccaccagca aggccgcggc cctgatgctg 86580agcgagtgtc tgcgcgcgga gttggcggag ttcggcatcg gtgtgagcgt ggtctgcccc 86640ggcctggtcc gtacgccgtt cgcgtccgcg atgtacttcg ccggcgcgtc ccccgacgag 86700cacacccggc tgcgtgagtc ctccgcccgc cgcttcgcgg gccgcggctg cccgccggag 86760aaggtcgcgg acgccgtcct gcgcgcgatc gtccgcaata cggcggtggt cgccgtcacc 86820cccgacgccc gcgccgtccg tctgatgagc cgcttcgcgc cccgcctccg cgccgtcgtg 86880gcccggctgg acccgtaggc agggcccgta cgggcagcgg gcgtccggtt cgggccaccg 86940gccgcggtat ccgcgcccct gcccggagct gtgccgctcc gggcaggggc gcgcggacga 87000ggcggtccgg cccggcggcc cggacctggc ggtccgttac tcaaaccgcg tgagcgtcag 87060ccggatcccg gtgggagcgg tgtcctggat gtaggaggcg aagtcggcca cgtcgtcgaa 87120ggcgaagccg taggctcggc cgtcctccgt gatcgcgtgc atcgccttgg cgtagtggtt 87180ggtcagcgcg gtcctgtaga aggccgcggg gtcggtcgtg ggctgggcgg cggaggtgag 87240cagggtcgag cggttgaatc cggcgccgag gaccgcggcg acgggaccgg tggtgccgtc 87300gttcggcgcg gcgagggcac cgtggcagaa gagcacgtcg cgcgtggtgg gcttggcgaa 87360ggacacctgg gcgggcccgt cgaaggtcag ccgctcgccg cgcacccggc cggtgaaggt 87420cccggcgttg gtggtgaccg tgaggtccct ggcggtgtag gtgctccaca cctcgtcgat 87480gtacggagcg aggtagtcct tcgggaacag gccggcgtcc agcccgtgcc cgggggcgat 87540cacacggagg tcgtccagga ccagtggcgc gaactccgcg acgcggcgga ccgcttcgaa 87600cgccgccgcc cggccgccgg cccgcacggt gccggtcgtc tggtccttcg cgcccgtcag 87660ccggatactg agcggcacgc tgaacatgtc caccatggtc gtgttgcaga acatgccgga 87720
ggggttgtag gtgaactcgg cgcagtcgtg cagcaccctg tagttcggat cggacgcgac 87780ccagccggcc gggtactgca gcgcggcgtt cccggctccg tccgtgacca ccttgaactt 87840gagtttctgt ccgagcgcga catagatccg gccggacatg tacggcaggg agagccgggt 87900ctcgccgctg ccggccagtg tgatcgcgta gtccgtgaag ccgtcggggc cgttgtccga 87960cagggcgacg ggcgcgaggg tgccgtcggg cgtgagccgt acctgtcggc cgtcctggtt 88020ccccacgacg tagacatgga cgtcgccgtt gccgaagacg ccggtgtcgt tgacgaccgt 88080cagcggcagg gcgcccgccg tggtcccttc cgcgtcgcgg tcccggccgg ggccggcgag 88140ggcgtgcggt gccagggctg cgacggcggg agcggccatc gcggcgccgc cgagggcgac 88200gagcagggtg cggcggccga ggctgcgctg gtgtcgagga gtcatgtggg gggcctcctg 88260gtgggcttgc cgatgttcta atgacgggaa catgacaggt gagaagcgtg ggagcgctcc 88320tcagggcccg atggtacgca cggggaggcg tcccgcgtcc ccgtgccggg accgcttaac 88380cgacgcttaa gggccgttta 88400<210>2<211>922<212>PRT<213>细菌<400>2Met Arg Gly Val Ser Pro Ser Val Ser Val Arg Glu Pro Gln Gly Leu1 5 10 15Thr Phe Leu Gly Leu Gly Arg Gln Ser His Ala Val Arg Thr Ala Leu20 25 30Glu Ala Cys Ala Ala Gly Arg Val Arg Val Leu Val Val Glu Gly Gly35 40 45
Leu Gly Cys Gly Lys Ser Ala Phe Leu Gly Glu Ala Leu Lys His Ala50 55 60Ala Ala Ser Gly Phe Leu Val Leu Arg Ser Ala Gly Ser Pro Pro Glu65 70 75 80Gly Arg Arg Pro Phe Asp Leu Leu Arg Gln Leu Ala Val Asp Pro Asp85 90 95Ile Pro Asp Ala Gln Arg Ser Leu Leu Gln Asp Ala Val Gly Thr Glu100 105 110Thr Pro Ala Ala Gln Arg Val Arg Ala Ala Leu His Gln Leu Thr Gly115 120 125Ala Ala Pro Val Val Ile Gly Ile Asp Asp Leu His His Ala Asp Pro130 135 140Gln Ser Leu His Cys Leu Leu Gln Ala Val Asp His Pro Arg Ala Thr145 150 155 160Arg Leu Leu Leu Val Cys Thr Ala Leu Pro Ser Gly Leu Ala Ala Asp165 170 175Pro Ala Val Glu Ala Glu Leu Leu Cys Gln Pro Ala Leu Gln Arg Val180 185 190Met Leu Gly Arg Leu Ser Leu Arg Ala Val Ser Gly Leu Arg Ala Ala195 200 205Arg Pro Gly Pro Ala Val Glu Ala Leu Pro Ala Asp Asp Leu Leu Ala210 215 220
Val Thr Gly Gly Asn Pro Leu Leu Val His Ala Leu Leu Glu Glu Leu225 230 235 240Val Glu Ser His Thr Gln Gly His Thr Asp Glu Arg Ala Gly Arg Arg245 250 255Arg Arg Ala Ala Ser Pro Val Ile Gly Gly Arg Phe Tyr Gln Ala Val260 265 270Leu Ala Ser Leu Ser Arg Thr Asp Ser Leu Val Arg His Ser Ala Gly275 280 285Ala Leu Ala Val Leu Gly Asp Ser Gly Cys Ala Glu Val Ile Ala Arg290 295 300Leu Leu Gly Ile Gly Arg Ala Met Ala Ala Arg Gly Leu Arg Ala Leu305 310 315 320Glu Ala Thr Gly Leu Thr Ala Ser Gly Arg Phe Arg His Pro Val Val325 330 335Glu Ala Ala Ala Leu Asp Thr Leu Asp His Asp His Arg Ala His Leu340 345 350His Arg Arg Ala Ala Ala Leu Leu Tyr Asp Val Gly Ala Glu Pro Asp355 360 365Glu Val Ala Arg His Leu Leu Ala Ala Arg His Ala Ala Gly Pro Trp370 375 380Ala Met Ser Val Leu Arg Asp Ala Ala Glu Gln Leu Leu Met Arg Asp385 390 395 400
Asp Val Leu Thr Ala Val Ser Cys Leu Glu Leu Ala Arg Arg Ser Cys405 410 415Ala Gly Gly Pro Arg Arg Ala Glu Ile Leu Leu Arg Leu Thr Val Ala420 425 430Thr Arg Arg Thr Asp Pro Ala Ala Ala Glu Asp His Leu Ala Glu Leu435 440 445Val Thr Glu Leu Arg Ala Gly Arg Leu Thr Ser Ala Glu Thr Glu Arg450 455 460Leu Gly His Leu Leu Leu Gly Cys Gly Arg Leu Glu Glu Ala Thr Glu465 470 475 480Val Met Gly Arg Pro Gly Pro His Gly Asp Pro Arg Thr Pro Arg Leu485 490 495Glu Thr Gly Phe His Ala Ser Ala Leu Trp Glu Pro Leu Ile Arg Pro500 505 510Arg Thr Asp Pro Glu Pro Gly Asp Glu Glu Ser Pro Arg Pro Arg Met515 520 525Pro Val Thr Gly Ile Trp Asp Leu Pro Gly Asp Gly Thr Asn Ala Ser530 535 540Ala Ser Asp Ala Ala Glu His Val Leu Arg Ser Leu Pro Leu Thr Asp545 550 555 560Thr Thr Leu Val Ile Val Val Asn Ala Val Arg Val Leu Cys Arg Thr565 570 575
Gly Ser Tyr Glu Thr Ala Ala Leu Trp Cys Thr Arg Leu Leu Gly Glu580 585 590Ala Ala Gly Arg Arg Leu Pro Gly Trp Lys Ala Gln Phe Leu Ala Leu595 600 605Gln Ala Glu Ile Ala Leu Cys Arg Gly Leu Leu Ala Asp Thr Glu Glu610 615 620Tyr Ala Arg Gln Ala Leu Ala Cys Val Pro Arg Cys Ser Arg Ser Val625 630 635 640Phe Ile Gly Gly Pro Leu Ala Ser Arg Val Phe Ala Ala Thr Ala Met645 650 655Gly Arg Tyr Asp Glu Ala Thr Arg Gln Leu Asp His Pro Val Pro Glu660 665 670Ala Leu Phe Arg Ser Val Tyr Gly Pro Ala Tyr Leu Arg Ala Arg Gly675 680 685His Tyr Tyr Leu Ala Leu Asp Arg Pro Leu Ala Ala Val Arg Asp Phe690 695 700Leu Gly Ala Gly Arg Leu Leu Arg Arg Trp Gly Ile Asp Arg Pro Thr705 710 715 720Leu Met Pro Trp Arg Ser Asp Ala Ala Glu Ala Phe Leu Arg Leu Cys725 730 735Glu Pro Arg Arg Ala Asp Arg Leu Leu Arg Glu Gln Leu Ala Arg Thr740 745 750
Pro Asp Asp Asp Pro His Val Arg Gly Val Ser Leu Arg Leu Arg Ala755 760 765Gln Ile Ala Glu Pro Pro Asp Arg Leu Asn Leu Leu Thr Glu Ala Val770 775 780Asn His Leu Lys Ser Ser Gly Asp Arg Leu Ala Leu Ala Gly Ala Leu785 790 795 800Ala Asp Leu Gly Ala Ala Tyr Arg Glu Arg Gly Glu Ser Thr Arg Ala805 810 815Gly Ala Thr Ile Arg Arg Ala Trp His Leu Ala Asn Asp Cys Gly Ala820 825 830Arg Ala Leu Cys Glu Arg Ile Leu Pro Gly Gly Pro Gly Arg Gln Ser835 840 845Phe Gly Asp Gly Thr Gly Arg Thr Glu Ala Ala Leu Ser Gly Ser Glu850 855 860Leu Arg Val Val Glu Leu Ala Ala Asn Gly His Thr Asn Arg Glu Ile865 870 875 880Ala Ala Arg Leu Cys Ile Thr Val Ser Thr Val Glu Gln His Leu Thr885 890 895Arg Ala Tyr Arg Lys Leu Glu Ile Ser Arg Arg Gln Glu Leu Pro Ala900 905 910Arg Leu Cys Ala His Ile Glu Ser Pro Val915 920
<210>3<211>259<212>PRT<213>细菌<400>3Met Pro Asp Leu Cys Glu Thr Glu Ser Leu Trp Leu Arg Arg Phe Gln1 5 10 15Pro Ala Pro Ala Ala Arg Thr Arg Leu Met Cys Phe Pro His Ala Gly20 25 30Gly Ser Ala Ser Ala Tyr Leu Arg Leu Ala Arg Ser Leu Ala Pro Gly35 40 45Ile Glu Val Leu Ala Val Gln Tyr Pro Gly Arg Gln Asp Arg Arg Ala50 55 60Glu Pro Cys Pro Asp Ser Val Glu Gly Leu Ala Asp Asp Leu Phe Ala65 70 75 80Ala Val Arg His Arg Val Asp Ala Ser Thr Ala Leu Phe Gly His Ser85 90 95Met Gly Ala Val Leu Ala Phe Glu Leu Ala Arg Arg Leu Glu Arg Asp100 105 110Ala Gly Val Arg Cys Ala Arg Ile Phe Ala Ser Gly Arg Arg Ala Pro115 120 125Ser Arg Phe Arg Asp Asp Ser Ala Pro Ala Ala Ser Asp Ala Ser Met130 135 140Leu Ala Glu Met Arg Thr Leu Gly Gly Thr Asp Leu Arg Val Leu Gln
145 150 155 160Asp Glu Glu Leu Leu Ile Ala Ala Leu Pro Ala Leu Arg Ala Asp Tyr165 170 175Arg Ala Ile Gly Thr Tyr Arg Ala Ala Asp Asp Ala Val Val Gly Cys180 185 190Pro Val Thr Val Leu Val Gly Asp Ala Asp Pro Arg Thr Ser Leu Asp195 200 205Asp Ala His Ala Trp Ser Ala His Thr Thr Ala Glu Ser Glu Val Leu210 215 220Thr Phe Ser Gly Gly His Phe Phe Leu Asp Ala His His Asp Ala Val225 230 235 240Val Glu Val Val Thr Ala Arg Leu Arg Gln Asp Arg Ala Pro Arg Pro245 250 255Asp Arg Val<210>4<211>267<212>PRT<213>细菌<400>4Met Pro Glu Leu Asn Asp Arg Thr Ala Leu Val Thr Gly Ala Ser Arg1 5 10 15Gly Ile Gly Lys Ala Ile Ala Gln Arg Leu Ala Ala Glu Gly Val Arg20 25 30
Val Ala Val His Tyr Gly Thr Gln Glu Lys Ser Ala Gln Glu Thr Val35 40 45Glu Thr Ile Glu Arg Ala Gly Gly Arg Ala Phe Ala Val Arg Ala Asp50 55 60Leu Leu Arg Asp Asp Ala Val Asp Glu Leu Phe Thr Ala Leu Glu Arg65 70 75 80Glu Leu Glu Gly Arg Pro Leu His Ile Leu Val Asn Asn Ala Ala Val85 90 95Ala Pro Ala Pro Gly Asp Pro Ala Leu Ala Ala Gln Asp Gly Tyr Val100 105 110Pro Gly Leu Ser Asp Thr Thr Pro Glu Glu Phe Asp Arg Val Tyr Arg115 120 125Ile Asn Val Arg Ala Pro Phe Phe Val Thr Gln Arg Ala Leu Ser Leu130 135 140Met Ala Asp Gly Gly Arg Ile Val Asn Val Ser Ser Ala Val Thr Arg145 150 155 160Ile Ala Trp Pro Leu Leu Pro Tyr Ala Met Thr Lys Gly Ala Leu Glu165 170 175Met Met Ala Pro Arg Leu Ala Asn Glu Leu Gly Ser Arg Gly Ile Thr180 185 190Val Asn Thr Val Ala Pro Gly Ile Thr Asp Thr Asp Met Asn Arg Trp195 200 205
Val Arg Glu Thr Pro Gly Ala Glu Ala Gly Ile Ser Ala Leu Thr Ala210 215 220Leu Gly Arg Leu Gly Arg Pro Asn Asp Ile Ala Gly Ile Val Ala Phe225 230 235 240Leu Val Ser Asp Asp Ala Arg Trp Ile Thr Gly Gln Leu Leu Asp Ala245 250 255Ser Gly Gly Met Ala Leu Ala Pro Ala Met Met260 265<210>5<211>2341<212>PRT<213>细菌<400>5Val His Glu Thr His Ala His Gly Glu Glu Gly Ser Ser Asp Gly Ser1 5 10 15Ala Asp Ala Val Val Phe Val Phe Pro Gly Gln Gly Ser Gln Trp Pro20 25 30Gly Met Gly Ala Glu Leu Trp Asp Thr Ser Pro Val Phe Arg Glu Ser35 40 45Val Arg Ala Cys Ala Asp Ala Leu Ala Pro Tyr Leu Asp Trp Ser Val50 55 60Glu Gly Val Leu Arg Gly Ala Pro Asp Ala Pro Ala Gly Pro Ala Leu65 70 75 80
Asp Arg Ala Asp Val Ala Gln Pro Ala Leu Phe Thr Leu Met Val Ser85 90 95Leu Ala Glu Leu Trp Arg Ser His Gly Val Glu Pro Cys Ala Val Leu100 105 110Gly His Ser Leu Gly Glu Ile Ala Ala Ala His Val Ala Gly Ala Leu115 120 125Thr Leu Ala Asp Ala Ala Arg Val Ala Ala Leu Trp Ser Arg Ala Gln130 135 140Ala Thr Leu Ser Gly Thr Gly Thr Leu Leu Ala Ala Lys Ala Ala Pro145 150 155 160Glu Glu Leu Ala Pro His Leu Gln Arg Trp Asn Gly Asp Asp Arg His165 170 175Gly Thr Arg Leu Ala Ile Ala Gly Val Asn Gly Pro Gly Ser Thr Val180 185 190Val Ala Gly Asp Leu Asp Ala Ile Ala Ala Leu Ala Ala Asp Leu Ala195 200 205Ser Ala Gly Val Arg Thr Arg Arg Val Ala Val Asp Val Pro Thr His210 215 220Ser Pro Ala Met Arg Thr Leu Arg Glu Arg Ile Leu Thr Asp Leu Ala225 230 235 240Ser Val Ala Pro Cys Val Ser Arg Leu Pro Phe His Ser Ser Leu Thr245 250 255
Gly Gly Leu Val Asp Thr Arg Gly Leu Asp Ala Asp Tyr Trp Tyr Arg260 265 270Asn Ile Ser Glu Thr Ala Arg Phe Asp Leu Ala Ala Arg Gly Leu Leu275 280 285Ala Asp Gly His Arg Thr Phe Val Glu Leu Ser Pro His Pro Ile Leu290 295 300Thr Leu Gly Leu Gln Ala Leu Ala Asp Asp Val Pro Gly Ala Ala Asp305 310 315 320Ala Leu Val Thr Gly Thr Leu Arg Arg Gly Arg Gly Gly Met Arg Gln325 330 335Phe Gln Asp Ala Leu Gly Arg Leu Ser Val Pro Ala Gly Gly Arg Pro340 345 350Gly Arg Glu Val Ser Ala Ala Ala Leu Ala Gly Arg Leu Ala Pro Leu355 360 365Ser Pro Ala Gln Gln Glu His Leu Leu Val Glu Leu Val Cys Ala His370 375 380Phe Ala Ala Leu Val Gly Gly Asp Gly Gly Ala Pro Pro Thr Val Arg385 390 395 400Pro Ser Ala Ala Phe Thr Asp Gln Gly Cys Asp Ser Ala Thr Ala Leu405 410 415Glu Leu Arg Asp Arg Leu Arg Glu Ala Thr Gly Leu Arg Leu Pro Ala420 425 430
Thr Leu Val Phe Asp His Pro Thr Pro Ala Ala Val Ala Gly Arg Leu435 440 445Arg Arg Leu Ala Leu Gly Ile Glu Glu Thr Ala Asp Thr Ala Pro Val450 455 460Ala Val Arg Gly His Arg Glu Gly Glu Pro Ile Ala Ile Val Gly Met465 470 475 480Ala Cys Arg Phe Pro Gly Gly Val Arg Ser Pro Glu Asp Leu Trp Arg485 490 495Leu Val Thr Glu Gly Gly Asp Ala Leu Gly Pro Phe Pro Thr Asp Arg500 505 510Gly Trp Asp Thr Gly Arg His Ala Glu Asp Pro Ala Thr Pro Gly Thr515 520 525Tyr Val Gln Gly Glu Gly Gly Phe Leu Tyr Asp Ala Gly Glu Phe Asp530 535 540Ala Glu Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro545 550 555 560Gln Gln Arg Leu Leu Leu Glu Met Ala Trp Glu Thr Phe Glu Arg Ala565 570 575Gly Ile Asp Pro Thr Ser Ala Arg Gly Ser Arg Thr Gly Val Phe Ala580 585 590Gly Val Leu Pro Leu Gly Tyr Gly Pro Arg Met Asp Glu Thr Asp Gln595 600 605
Gly Thr Ala Asp Leu Gln Gly His Leu Leu Thr Gly Thr Leu Pro Ser610 615 620Val Ala Ser Gly Arg Ile Ser Tyr Thr Leu Gly Leu Glu Gly Pro Ala625 630 635 640Val Ser Val Glu Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu645 650 655Ala Cys Arg Ser Leu Arg Ala Gly Glu Cys Asp Leu Ala Leu Thr Gly660 665 670Gly Val Ser Val Leu Ala Thr Leu Gly Leu Phe Val Glu Phe Ser Arg675 680 685Gln Arg Gly Leu Ser Ala Asp Gly Arg Cys Lys Ala Tyr Ala Ala Ala690 695 700Ala Asp Gly Thr Gly Trp Ser Glu Gly Ala Gly Leu Leu Leu Val Glu705 710 715 720Arg Leu Ser Asp Ala Arg Arg Leu Gly His Arg Val Leu Ala Val Val725 730 735Arg Gly Ser Ala Ile Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala740 745 750Pro Ser Gly Pro Ser Gln Gln Arg Val Ile Arg Glu Ala Leu Ala Asp755 760 765Ala Gly Leu Thr Ala Ala Asp Val Asp Ala Val Glu Gly His Gly Thr770 775 780
Gly Thr Arg Leu Gly Asp Pro Ile Glu Ile Glu Ala Leu Leu Ala Thr785 790 795 800Tyr Gly Gln Gly Arg Ala Arg Glu Arg Pro Leu Trp Leu Gly Ser Leu805 810 815Lys Ser Asn Ile Gly His Thr Met Ala Ala Ala Gly Val Gly Gly Val820 825 830Ile Lys Met Val Met Ala Leu Arg His Gly Glu Leu Pro Arg Thr Leu835 840 845His Val Asp Ala Pro Ser Pro Arg Ala Asp Trp Ser Ala Gly Glu Val850 855 860Arg Leu Leu Thr Glu Ala Val Ala Trp Pro Ala Ala Ala Asp Gly Glu865 870 875 880Pro Arg Arg Ala Gly Val Ser Ser Phe Gly Val Ser Gly Thr Asn Ala885 890 895His Ala Ile Leu Glu Glu Ala Pro Ala Pro Glu Asp Glu Glu Pro Ala900 905 910Pro Pro Asp Gly Glu Ala Leu Leu Pro Trp Ala Val Ser Thr Arg Ser915 920 925Glu Ala Ala Leu Arg Thr Gln Ala Arg Met Leu Ala Asp Val Val Arg930 935 940Asp Asp Pro Gly Val Gly Leu Ala Asp Val Gly Ala Glu Leu Ala Arg945 950 955 960
Gly Arg Ala Ala Leu Glu His Arg Ala Val Val Ile Ala Ser Gly Arg965 970 975Ala Glu Phe Ala Arg Ala Leu Glu Ala Val Ala Ser Gly Glu Pro His980 985 990Pro Ala Val Val Arg Gly His Ala Gly Ser Glu Arg Gly Gly Val Val995 1000 1005Phe Val Phe Pro Gly Gln Gly Gly Gln Trp Ala Gly Met Gly Leu1010 1015 1020Asp Leu Leu Arg Ser Ser Pro Val Phe Ala Glu His Ile Ala Ala1025 1030 1035Cys Gly Lys Ala Leu Ala Pro Trp Val Lys Trp Ser Leu Thr Glu1040 1045 1050Val Leu His Arg Asp Ala Glu Asp Pro Val Trp Asp Arg Ala Asp1055 1060 1065Val Val Gln Pro Val Leu Phe Ser Val Met Thr Ser Leu Ala Ala1070 1075 1080Leu Trp Arg Ser Tyr Gly Val Glu Pro Asp Ala Val Thr Gly His1085 1090 1095Ser Gln Gly Glu Ile Ala Ala Ala Tyr Val Cys Gly Ala Leu Gly1100 1105 1110Leu Glu Asp Ala Ala Arg Thr Val Ala Leu Arg Ser Arg Ala Leu1115 1120 1125
Val Ala Leu Arg Gly Arg Gly Gly Met Ala Ser Val Ala Ser Ala1130 1135 1140Ala Pro Asp Val Glu Glu Leu Ile Ala Arg Arg Trp Pro Gly Arg1145 1150 1155Leu Trp Val Ala Ala Phe Asn Gly Pro Gly Ala Val Thr Val Ser1160 1165 1170Gly Asp Gly Asp Ala Leu Glu Glu Phe Leu Gly His Cys Ala Asp1175 1180 1185Thr Glu Val Arg Ala Arg Arg Val Pro Val Asp Tyr Ala Ser His1190 1195 1200Cys Pro His Thr Glu Ala Ile Glu Arg Glu Leu Leu Asp Ala Leu1205 1210 1215Glu Asp Ile Thr Pro Arg Pro Ala Ala Val Pro Phe Tyr Ser Thr1220 1225 1230Val Asp Asp Ala Trp Leu Asp Thr Thr Arg Leu Asp Ala Ser Tyr1235 1240 1245Trp Tyr Arg Asn Leu Arg Arg Pro Val Arg Phe Ser Gln Ala Val1250 1255 1260Arg Ala Leu Thr Asp Gly Gly His Arg Val Phe Ile Glu Ala Ser1265 1270 1275Pro His Pro Thr Leu Val Pro Ala Ile Glu Asp His Gly Asp Val1280 1285 1290
Thr Ala Leu Gly Thr Leu Arg Arg His Gly Asp Asp Thr Glu Arg1295 1300 1305Phe Leu Thr Ala Leu Ala His Leu His Val Thr Gly Ala Ala Gly1310 1315 1320Gln Asp Leu Trp Arg His His Tyr Ala Arg Leu Arg Pro Ala Pro1325 1330 1335Arg His Val Asp Leu Pro Thr Tyr Ala Phe Gln Arg Asp Arg Tyr1340 1345 1350Trp Trp Ser Gly Gly Ala Gly Arg Gly Asp Val Thr Thr Ala Gly1355 1360 1365Leu His Pro Gly Gly His Pro Leu Leu Gly Ala Ala Leu Asp Leu1370 1375 1380Ala Asp Gly Gly Gly Arg Leu His Thr Gly Arg Val Ser Leu Arg1385 1390 1395Thr His Pro Trp Ile Ala Asp His Gly Val Ala Gly Ile Thr Leu1400 1405 1410Leu Pro Gly Thr Ala Phe Leu Glu Leu Ala Leu His Thr Gly Glu1415 1420 1425Ser Gly Asn Val Arg Glu Leu Thr Leu His Ala Pro Leu Val Val1430 1435 1440Pro Asp Glu Glu Gly Val Asp Leu Gln Val His Leu Ala Arg Pro1445 1450 1455
Asp Glu Ala Gly Leu Arg Ala Leu Thr Arg Leu Leu Pro Gly Arg1460 1465 1470Gly Val Pro Thr Pro Arg Ala Pro Trp Gln Pro His Ala Thr Gly1475 1480 1485Leu Leu Gly Pro Ala Asp Arg Ala Pro Gly Ser Ser Gly Leu Glu1490 1495 1500Pro His Asp Leu Gly Gly Ala Trp Pro Pro Pro Gly Ala Val Pro1505 1510 1515Leu Val Pro Gly Glu Leu Gly Asp Val Pro Gly Cys Tyr Ala Arg1520 1525 1530Leu Ala Asp Glu Gly Phe Glu Tyr Gly Pro Ala Phe Arg Gly Leu1535 1540 1545Arg Ala Val Trp Arg Arg Gly Thr Glu Ile Phe Ala Glu Val Ala1550 1555 1560Leu Pro Ala Gly Asp Gly Ser Val Phe Arg Leu His Pro Ala Leu1565 1570 1575Leu Asp Ala Val Leu His Pro Val Val Leu Gly Leu Val Asp Gly1580 1585 1590Val Pro Ala Arg Pro Leu Pro Phe Ser Trp Asn Gly Val Ala Leu1595 1600 1605His Ala Pro Ala Ser Gly Ala Leu Arg Val Arg Leu Ala Pro Ala1610 1615 1620
Asp Asp Gly Ala Val Gly Ile Thr Ala Ala Thr Ala Ala Gly Glu1625 1630 1635Pro Val Leu Ser Val Ala Ala Leu Ala Leu Arg Ser Ala Ser Ala1640 1645 1650Glu Gln Leu Arg Ala Ala Ile Arg Ser Ala Ala Gly Ser Arg Asp1655 1660 1665Ala Leu Tyr Glu Leu Asp Trp Leu Pro Leu Pro Ala Asp Arg Ala1670 1675 1680Ala Ser Pro Gly Gly Ala Asp Ile Ala Ala Leu Gly Thr Ser Glu1685 1690 1695Leu Pro Cys Arg Thr Tyr Glu Thr Ile Ala Glu Leu Ser Gln Ala1700 1705 1710Leu Ala Asp Gly Ala Pro Ala Pro Asp Ala Val Val Ser Asp Val1715 1720 1725Gly Ala Val Gly Gly Pro Leu Asp Thr Val Ser Leu His Gly Leu1730 1735 1740Cys Arg Arg Gly Leu Glu Leu Val Gln Ala Trp Leu Gly Glu Pro1745 1750 1755Arg Thr Ala Asp Thr Arg Leu Val Leu Val Thr Arg Gly Ala Val1760 1765 1770Gly Cys Ala Pro Ala Glu Pro Val Ala Asp Pro Ala Ala Ala Ala1775 1780 1785
Leu Trp Gly Leu Val Arg Ser Ala Gln Ala Glu His Pro Gly Arg1790 1795 1800Leu Leu Leu Leu Asp Leu Asp Pro Ala Gly Ser Arg Pro Val Ser1805 1810 1815Gly Arg Leu Val Glu Gln Ala Val Ala Cys Gly Glu Pro His Ile1820 1825 1830Ala Val Arg Gly Asp Gly Leu Arg Val Pro Arg Leu Ser Arg Ala1835 1840 1845Thr Ala Ala Pro Ala His Pro Pro Ala Gly Gly Arg Glu Ala Gln1850 1855 1860Trp Asp Pro Glu Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Ser1865 1870 1875Leu Gly Ala Leu Phe Ala Arg His Leu Val Thr Ala His Gly Val1880 1885 1890Arg Arg Leu Leu Leu Ala Ser Arg Ser Gly Pro Gly Ala Pro Gly1895 1900 1905Ala Ala Gly Leu Arg Asp Glu Leu Thr Ala His Gly Ala Thr Val1910 1915 1920Thr Val Ala Ala Cys Asp Val Ala Asp Arg Glu Ala Val Ala Ala1925 1930 1935Leu Leu Ala Ser Val Pro Ser Glu His Pro Leu Thr Ala Val Val1940 1945 1950
His Thr Ala Gly Val Leu Asp Asp Gly Val Leu Ala Ser Leu Thr1955 1960 1965Ala Asp Arg Leu Ala Arg Val Leu Arg Ala Lys Ala Asp Ala Ala1970 1975 1980Leu His Leu His Asp Leu Thr Arg Asp Leu Pro Leu Ala Ala Phe1985 1990 1995Val Leu Phe Ser Ser Val Thr Ala Thr Leu Gly Thr Pro Gly Gln2000 2005 2010Ala Asn Tyr Thr Ala Ala Asn Ala Phe Leu Asp Ala Leu Ala Arg2015 2020 2025His Arg Arg Ala Ala Gly Leu Pro Ala Val Ser Leu Ala Trp Gly2030 2035 2040Leu Trp Glu Gln Thr Gly Gly Leu Thr Asp His Leu Gly Ser Val2045 2050 2055Asp Leu Arg Arg Met Ala Arg Asn Gly Leu Val Ala Leu Pro Ala2060 2065 2070Asp Ala Gly Leu Ala Leu Phe Asp Thr Ala Leu Ala Leu Asp Arg2075 2080 2085Ala Asn Leu Val Pro Ala Arg Leu Asp Leu Pro Ala Leu Arg Arg2090 2095 2100Ala Thr His Val Pro Pro Val Leu Arg Arg Leu Val Glu Val Pro2105 2110 2115
Gly Ala Pro Ser Ala Asp Arg Ser Ala Gly Ser Gly Gly Glu Val2120 2125 2130Arg Pro Leu Arg Glu Thr Leu Ala Gly Leu Asp Asp Arg Lys Arg2135 2140 2145Pro Ala Ala Val Ser Arg Leu Val Arg Arg His Val Ala Trp Val2150 2155 2160Leu Gly Ala Asp Gly Pro Glu Ser Val Asp Glu Asp Arg Ser Phe2165 2170 2175Arg Asp Leu Gly Phe Asp Ser Leu Met Ala Val Glu Leu Arg Asn2180 2185 2190Gln Leu Asn Thr Ala Ala Gly Ile Arg Leu Ala Ala Thr Leu Val2195 2200 2205Phe Asp His Pro Thr Pro Ser Ala Val Ala Arg His Leu Leu Asp2210 2215 2220Arg Cys Ser Pro Asp Pro Ala Ala Pro Ala Ala Pro Ser Gly Thr2225 2230 2235Ala Val Ala Ser Ala Leu Ala Thr Leu Ala Glu Leu Glu Thr Ala2240 2245 2250Leu Asn Gly Ile Pro Ala Glu Glu Trp Thr Ala Ala Gly Gly Pro2255 2260 2265Ala Arg Leu Met Thr Leu Ala Ser Ser Leu Pro Ala Pro Ala Ser2270 2275 2280
Val Pro Arg Thr Pro Ala Ala Gly Glu Ala Ala Glu Lys Leu Ala2285 2290 2295His Ala Ser Arg Asp Glu Ile Phe Ala Phe Ile Asp Arg Glu Leu2300 2305 2310Gly Arg Asp Ser Gly Pro Ala Ser Pro Ser Arg Leu Gly Pro Gln2315 2320 2325Thr Pro Asp Ser Thr Asp Lys Ala Pro Phe His Gly Glu2330 2335 2340<210>6<211>3723<212>PRT<213>细菌<400>6Met Glu Asn Glu Glu Lys Leu Leu Asp Tyr Leu Lys Trp Val Thr Ala1 5 10 15Asp Leu His Arg Ser Arg Glu Arg Val Thr Glu Leu Glu Glu Ala Gly20 25 30Arg Glu Pro Ile Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly Glu35 40 45Val Arg Ser Pro Glu Glu Leu Trp Gly Leu Val Ala Ser Gly Gly Asp50 55 60Ala lle Gly Ala Phe Pro Asp Asp Arg Gly Trp Asp Leu Asp Gly Leu65 70 75 80
Phe Asp Pro Asp Pro Glu Arg Ala Gly Thr Ser Tyr Thr Arg Arg Gly85 90 95Gly Phe Leu Tyr Asp Ala Ala Glu Phe Asp Ala Gly Phe Phe Gly Ile100 105 110Ser Pro Arg Glu Ala Met Ala Met Asp Pro Gln Gln Arg Leu Leu Leu115 120 125Glu Thr Ser Trp Glu Ala Phe Glu Arg Ala Gly Ile Asp Pro Ser Ser130 135 140Val Arg Gly Ser Arg Val Gly Val Phe Ala Gly Leu Met Tyr His Asp145 150 155 160Tyr Ala Ala Ala Gln Gly Ser Thr Gly Asp Gly Asp Gly Glu Pro Asp165 170 175Phe Glu Gly Tyr Leu Gly Asp Gly Ser Val Ser Ser lle Ala Ser Gly180 185 190Arg Ile Ala Tyr Thr Leu Gly Leu Ala Gly Ala Ala Ile Thr Val Asp195 200 205Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala210 215 220Leu Arg Thr Gly Asp Ser Glu Leu Ala Leu Ala Gly Gly Val Ser Val225 230 235 240Met Ser Thr Pro Arg Thr Phe Val Gln Phe Ser Arg Gln Arg Gly Leu245 250 255
Ser Ala Asp Gly Arg Cys Lys Ala Tyr Ala Ala Ala Ala Asp Gly Thr260 265 270Gly Phe Ser Glu Gly Val Gly Met Val Leu Val Glu Arg Leu Ser Asp275 280 285Ala Arg Arg Leu Gly His Pro Val Leu Ala Val Val Arg Gly Ser Ala290 295 300Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro305 310 315 320Ser Gln Glu Arg Val Ile Arg Glu Ala Leu Ala Asn Ala Gly Leu Thr325 330 335Ala Ala Asp Val Asp Ala Val Glu Gly His Gly Thr Gly Thr Arg Leu340 345 350Gly Asp Pro Ile Glu Leu Gln Ala Leu Leu Ala Thr Tyr Gly Gln Gly355 360 365Arg Ala Arg Glu Arg Pro Leu Trp Leu Gly Ser Val Lys Ser Asn Ile370 375 380Gly His Ala Gln Ala Ala Ala Gly Val Gly Gly Val Ile Lys Met Val385 390 395 400Met Ala Leu Arg His Gly Glu Leu Pro Arg Thr Leu His Val Asp Ala405 410 415Pro Ser Pro Arg Val Asp Trp Ser Ala Gly Glu Val Arg Leu Leu Thr420 425 430
Glu Ala Val Ala Trp Pro Ala Ala Ala Asp Gly Glu Pro Arg Arg Ala435 440 445Gly Val Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Val Ile Leu450 455 460Glu Glu Ala Pro Ala Ser Glu Gly Glu Glu Ala Pro Pro Pro Glu Pro465 470 475 480Gly Ser Pro Leu Pro Trp Val Val Ser Gly His Ser Glu Ala Gly Leu485 490 495Arg Ala Gln Ala Gln Ala Leu Ala Glu Phe Ala Arg Thr Ala Pro Gly500 505 510Ala Glu Leu Val Asp Val Gly Ala Ala Leu Ala Arg Gly Arg Ala Ala515 520 525Leu Gly His Arg Ala Val Val Val Ala Ser Glu Arg Glu Glu Phe Glu530 535 540Arg Ala Leu Ala Ala Leu Ala Cys Gly Glu Pro His Pro Cys Val Val545 550 555 560Asp Gly Ser Ala Asp Gly Arg Arg Glu Asp Gly Val Val Phe Val Phe565 570 575Pro Gly Gln Gly Gly Gln Trp Ala Gly Met Gly Leu Asp Leu Leu Thr580 585 590Thr Ser Gly Val Phe Ala Glu His Ile Gly Ala Cys Glu Arg Ala Leu595 600 605
Ala Pro Trp Val Glu Trp Ser Leu Thr Glu Met Leu His Arg Glu Ala610 615 620Glu Asp Pro Val Trp Glu Arg Ala Asp Ile Val Gln Pro Val Leu Phe625 630 635 640Ser Val Met Val Ser Leu Ala Ala Leu Trp Arg Ser Tyr Gly Ile Glu645 650 655Pro Asp Ala Val Val Gly His Ser Gln Gly Glu Ile Ala Ala Ala His660 665 670Val Cys Gly Ala Leu Thr Leu Glu Asp Ala Ala Lys Val Val Ala Leu675 680 685Arg Ser Arg Ala Leu Ala Ala Leu Arg Gly Arg Gly Gly Met Val Ser690 695 700Leu Ser Leu Ser Thr Ala Asp Ala Gly Glu Leu Val Glu Arg Arg Trp705 710 715 720Ala Gly Arg Leu Trp Val Ala Ala Leu Asn Gly Pro Glu Ala Thr Thr725 730 735Val Ser Gly Asp Val Asp Ala Leu Glu Glu Leu Leu Ala His Cys Ala740 745 750Lys Ser Glu Val Arg Ala Arg Arg Val Pro Val Asp Tyr Ala Ser His755 760 765Cys Pro His Thr Glu Ala Ile Ala Glu Glu Ile Val Asp Ser Leu Gly770 775 780
Asp Ile Thr Pro Arg Ala Ala Thr Val Pro Phe Tyr Ser Thr Val Asp785 790 795 800Asp Met Trp Leu Asp Thr Thr Arg Leu Asp Ala Ser Tyr Trp Tyr Arg805 810 815Asn Leu Arg Leu Pro Val Arg Phe Ser Gln Ala Val Arg Ala Leu Thr820 825 830Glu Glu Gly His Arg Leu Phe Ile Glu Thr Ser Pro His Pro Thr Leu835 840 845Val Pro Ala Ile Glu Asp His Gly Asp Val Thr Ala Leu Gly Thr Leu850 855 860Arg Arg His Gly Asp Asp Thr Glu Arg Phe Leu Thr Ala Leu Ala His865 870 875 880Leu His Val Thr Gly Ala Ala Gly Gln Asp Leu Trp Arg His His Tyr885 890 895Ala Arg Leu Arg Pro Ala Pro Arg His Val Asp Leu Pro Thr Tyr Pro900 905 910Phe Gln Arg Arg Arg Tyr Trp Leu Glu Lys Pro Asp Pro Gln Thr Arg915 920 925Pro Gln Arg Ser Arg Ser Thr Ala Pro Asp Leu Asp Arg Leu Glu Ala930 935 940Glu Phe Trp Gln Ala Val Glu Glu Thr Asp Thr Asp Thr Leu Ala His945 950 955 960
Thr Leu His Leu Asp Thr Gln Thr Leu Glu Pro Val Leu Pro Ala Leu965 970 975Ala Thr Trp His Gln Gln Gln Arg Asp His Ala Arg Ile Asn Thr Trp980 985 990Thr Tyr Gln Glu Thr Trp Lys Pro Leu His Leu Pro Thr Thr Arg Pro995 1000 1005Thr Thr Pro Thr Ser Trp Leu Ile Ala Ile Pro Glu Thr His Arg1010 1015 1020Asn His Pro His Thr Thr Asn Leu Leu Thr Asn Leu Pro His His1025 1030 1035Asn Ile Thr Pro Ile Pro Leu Thr Ile Asn His Thr Thr Asp Leu1040 1045 1050His His Ala Tyr His His Ala His His His Thr Thr Pro Pro Ile1055 1060 1065Thr Ala Val Leu Ser Leu Leu Ala Leu Asp Glu Thr Pro His Pro1070 1075 1080His His Pro His Thr Pro Thr Gly Thr Leu Leu Asn Leu Thr Leu1085 1090 1095Thr Gln Thr His Thr Gln Thr His Pro Pro Thr Pro Leu Trp Tyr1100 1105 1110Leu Thr Thr Gln Ala Thr Thr Thr His Pro Asn Asp Pro Leu Thr1115 1120 1125
His Pro Thr Gln Ala Gln Thr Ile Gly Leu Ala Arg Thr Thr His1130 1135 1140Leu Glu His Pro His His Thr Gly Gly His Ile Asp Leu Pro Thr1145 1150 1155Thr Pro His Pro Asn Thr Leu Thr Gln Leu Ile Thr Ala Leu Thr1160 1165 1170His Pro His His Gln His Asn Leu Thr Ile Arg Thr His Thr Thr1175 1180 1185His Thr Arg Arg Leu Thr Pro Thr Thr Leu Gln Pro Thr Thr Pro1190 1195 1200Thr Pro Pro Thr Asn Pro His Gly Thr Thr Leu Ile Thr Gly Gly1205 1210 1215Thr Gly Ala Leu Ala Thr Thr Leu Ala His His Leu Ala Thr Thr1220 1225 1230Gly Thr Gln His Leu Leu Leu Thr Ser Arg Arg Gly Pro His Thr1235 1240 1245Pro Gly Ala Arg Gln Leu His Thr Gln Leu Thr Gln Leu Gly Thr1250 1255 1260Asn Thr Thr Ile Thr Ala Cys Asp Leu Ser Asp Pro Asp Gln Leu1265 1270 1275Thr His Leu Leu Thr His Ile Pro Pro Glu His Pro Leu Thr Thr1280 1285 1290
Val Ile His Thr Ala Gly Ile Leu Asp Asp Ala Thr Leu Thr Asn1295 1300 1305Leu Thr Pro Thr Gln Leu Asp Asn Val Leu Arg Ala Lys Ala His1310 1315 1320Thr Ala His Leu Leu His His Ala Thr Leu His Thr Pro Leu Asp1325 1330 1335His Phe Val Leu Tyr Ser Ser Ala Ala Ala Thr Leu Gly Ala Pro1340 1345 1350Gly Gln Ala Asn Tyr Ala Ala Ala Asn Ala Tyr Leu Asp Ala Leu1355 1360 1365Ala His His Arg His Thr His Asn Leu Pro Ala Thr Thr Ile Ala1370 1375 1380Trp Gly Thr Trp Gln Gly Asn Gly Leu Ala Asp Ser Asp Lys Ala1385 1390 1395Arg Ala Asn Leu Asp Arg Arg Gly Phe Leu Pro Met Pro Glu Thr1400 1405 1410Leu Ala Ala Ala Ala Ala Val Arg Ala Ile Glu Ser Arg Arg Pro1415 1420 1425Ser Val Val Ile Ala Ala Ile Asp Trp Ala Arg Ala Glu Arg Thr1430 1435 1440Pro Asp Val Glu Asp Leu Leu Pro Ala Ala Asp Glu Gly Ser Ser1445 1450 1455
Ser Gly Lys Pro Glu Ala Ala Pro Val Asp Leu Arg Gly Thr Leu1460 1465 1470Ser Arg Gln Ser Ala Ala Asp Gln Gln Ala Thr Leu Leu Gly Leu1475 1480 1485Val Arg Thr Gln Ala Ala Val Val Leu Arg His Thr Glu Pro Glu1490 1495 1500Ala Leu Ala Pro Gly Gln Ala Phe Arg Ala Leu Gly Phe Asp Ser1505 1510 1515Leu Thr Ala Val Glu Leu Arg Asn Arg Leu Ala Lys Ala Thr Asp1520 1525 1530Leu Ala Leu Pro Ala Ser Leu Val Phe Asp His Pro Thr Pro Val1535 1540 1545Lys Leu Ala Glu Phe Leu Arg Thr Glu Leu Leu Gly Thr Ala Pro1550 1555 1560Ala Thr Thr Ala Ala Val Pro Ala Leu Gln Ala His Thr Asp Glu1565 1570 1575Pro Ile Ala Ile Ile Gly Met Ala Cys Arg Phe Pro Gly Ala Val1580 1585 1590Thr Thr Pro Glu His Leu Trp Asn Leu Ile Ala Thr Glu Gln Asp1595 1600 1605Ala Ile Gly Glu Phe Pro Thr Asp Arg Gly Trp Asp Leu Asp Asn1610 1615 1620
Leu Tyr His Pro Asp Pro Asp His Pro Gly Thr Thr Tyr Thr Arg1625 1630 1635His Gly Gly Phe Leu His Asp Ala Gly Asp Phe Asp Ala Asp Phe1640 1645 1650Phe Gly Ile Asn Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln1655 1660 1665Arg Leu Leu Leu Glu Thr Ala Trp Glu Ala Ile Glu His Ala Gly1670 1675 1680Ile Leu Pro Asp Ala Leu His Gly Thr Pro Thr Gly Val Phe Thr1685 1690 1695Gly Val Asn Ala Gln Asp Tyr Ala Ala His Thr His Thr Ser Pro1700 1705 1710His Thr Thr Glu Gly Tyr Thr Leu Thr Gly Thr Ala Gly Ser Ile1715 1720 1725Ala Ser Gly Arg Ile Ala Tyr Val Leu Gly Leu Glu Gly Pro Ala1730 1735 1740Val Thr Ile Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His1745 1750 1755Leu Ala Cys Gln Ala Leu Arg Ala Gly Glu Cys Thr Thr Ala Leu1760 1765 1770Ala Ser Gly Ile Ser Ile Met Thr Thr Pro Leu Ala Phe Thr Glu1775 1780 1785
Phe Ser Arg Gln Arg Gly Leu Ala Ala Asp Gly Arg Cys Lys Ala1790 1795 1800Phe Ala Ala Ala Ala Asp Gly Thr Gly Trp Ser Glu Gly Val Gly1805 1810 1815Thr Leu Leu Leu Glu Arg Leu Ser Asp Ala Glu Arg Asn Gly His1820 1825 1830Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly1835 1840 1845Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Gln Arg1850 1855 1860Val Ile Arg Gln Ala Leu Val Asn Ala Asn Leu Ser Ala Val Asp1865 1870 1875Val Asp Ala Val Glu Ala His Gly Thr Gly Thr Lys Leu Gly Asp1880 1885 1890Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Gly Arg1895 1900 1905Ala Gln Glu Gln Pro Leu Trp Leu Gly Ser Val Lys Ser Asn Leu1910 1915 1920Gly His Thr Gln Ala Ala Ala Gly Met Ala Gly Leu Ile Lys Met1925 1930 1935Val Met Ala Leu Arg His Glu Ser Leu Pro Arg Thr Leu His Val1940 1945 1950
Asp Glu Pro Ser Pro Glu Val Asp Trp Ser Ser Gly Ala Val Ser1955 1960 1965Leu Leu Thr Glu Ala Arg Pro Trp Pro Arg Val Glu Asp Arg Pro1970 1975 1980Arg Arg Ala Gly Val Ser Ser Phe Gly Val Ser Gly Thr Asn Ala1985 1990 1995His Val Ile Val Glu Glu Ala Pro Ala Pro Thr Gly Val Glu Ala2000 2005 2010Val Glu Ala Ala Pro Ala Gly Val Glu Thr Ala Ala Ala Ala Ala2015 2020 2025Val Val Val Glu Thr Asp Gly Ala Gly Arg Val Ser Ala Asp Leu2030 2035 2040Pro Leu Val Trp Val Ala Ser Gly Lys Ser Gln Ala Ala Ile Arg2045 2050 2055Ala Gln Ala Ala Ala Leu His Ala His Val Leu Asp His Pro Glu2060 2065 2070Gln Asp Ala Asp Asp Ile Gly Tyr Ser Leu Ala Thr Thr Arg Ala2075 2080 2085Leu Phe Asp His Arg Ala Thr Leu Ile Ala Pro Asp Arg His Thr2090 2095 2100Val Pro Glu Pro Leu Thr Gly Leu Gly Asp Gly Arg Thr His Pro2105 2110 2115
His Leu Ile Pro Thr Pro Pro Thr Glu Pro Gly His Thr His Lys2120 2125 2130Ile Ala Phe Leu Cys Ser Gly Gln Gly Thr Gln Arg Pro Gly Met2135 2140 2145Ala Thr Gly Leu Tyr His Thr Tyr Pro Ala Phe Ala Ala Ala Leu2150 2155 2160Asp Glu Thr Cys Ala His Phe Asp Pro His Leu Asp His Pro Leu2165 2170 2175His Asp Leu Leu Leu Asn His Asp Pro Thr Asp Leu Leu Thr His2180 2185 2190Thr Leu Tyr Ala Gln Pro Ala Leu Phe Thr Leu Gln Lys Ala Leu2195 2200 2205His His Leu Ile Thr Glu Thr Tyr Gly Ile Thr Pro His Tyr Leu2210 2215 2220Ala Gly His Ser Leu Gly Glu Ile Thr Ala Ala His Leu Ala Gly2225 2230 2235Ile Leu Thr Leu Pro Asp Ala Thr His Leu Ile Thr Thr Arg Ala2240 2245 2250Arg Leu Met Gln Thr Met Pro Pro Gly Thr Met Thr Thr Leu His2255 2260 2265Thr Thr Pro Glu His Ile Gln Pro Leu Leu Asp Gln His Pro Gly2270 2275 2280
Lys Ala Ala Ile Ala Ala Val Asn Ser Pro His Ser Leu Val Ile2285 2290 2295Ser Gly Asp Pro Asp Thr Ile His His Ile Thr Thr Thr Cys His2300 2305 2310Asn Gln Gly Ile Thr Thr Lys Pro Leu Ala Thr Asn His Ala Phe2315 2320 2325His Ser Pro His Thr Asp Thr Ile Leu Glu Gln Leu Asp Thr Thr2330 2335 2340Thr His Thr Leu Thr Tyr His Gln Pro His Thr Pro Leu Ile Thr2345 2350 2355Ser Thr Pro Gly Asp Pro Leu Thr Pro His Tyr Trp Thr His Gln2360 2365 2370Thr Arg Gln Pro Val His Trp Thr Asp Thr Ile His Thr Leu His2375 2380 2385Thr His Gly Val Thr Thr Tyr Ile Ala Leu Gly Pro Glu His Thr2390 2395 2400Leu Thr Thr Leu Thr His His Asn Val Pro His His Gln Pro Thr2405 2410 2415Ala Ile Thr Leu Thr His Pro His His Asn Pro Thr His His Leu2420 2425 2430Leu Thr Ala Leu Ala His Leu His Thr Thr Gln Pro Thr Gly Pro2435 2440 2445
Asn Ile Trp His His His Tyr Thr Pro Val Ala Pro Ala Pro Arg2450 2455 2460His Val Asp Leu Pro Thr Tyr Pro Phe Pro Arg Arg Arg Tyr Trp2465 2470 2475Val Gln Ala Ser Ala Gly Thr Gly Asp Val Ser Ala Ala Gly Leu2480 2485 2490Gln Arg Pro Asp His Pro Leu Leu Gly Ala Val Met Glu Leu Ala2495 2500 2505Asp Gly Asp Gly Ile Val Leu Thr Gly Arg Leu Ser Leu His Thr2510 2515 2520His Pro Trp Leu Ala Asp His Ser Val Gly Gly Val Ala Leu Leu2525 2530 2535Pro Gly Thr Ala Leu Leu Glu Leu Ala Phe Gln Ala Gly Leu Arg2540 2545 2550Ala Gly Cys Pro Gly Val Asp Glu Leu Thr Leu His Ala Pro Leu2555 2560 2565Val Val Pro Glu Ser Gly His Val Val Val Gln Val Ser Val Ser2570 2575 2580Val Pro Gly Glu Ala Gly Arg Arg Gly Val Ser Val Tyr Gly Arg2585 2590 2595Leu Val Glu Asp Gly Gly Leu Glu Gly Glu Trp Thr Arg His Ala2600 2605 2610
Glu Gly Val Val Cys Pro Ser Val Pro Gly Glu Ser Val Val Val2615 2620 2625Glu Pro Val Ala Asp Gly Val Trp Pro Pro Ser Gly Ala Gln Pro2630 2635 2640Val Asp Leu Glu Glu Phe Tyr Gly Arg Leu Ala Gly Gly Gly Phe2645 2650 2655Val Tyr Gly Pro Val Phe Gln Gly Leu Cys Ala Ala Trp Arg Asp2660 2665 2670Gly Asp Asp Val Val Ala Glu Val Arg Leu Pro Asp Glu Gly Leu2675 2680 2685Ala Asp Val Ala Gly Phe Gly Val His Pro Ala Leu Leu Asp Ala2690 2695 2700Ala Val Gln Ala Val Thr Leu Leu Phe Pro Asp Gln Gln Gln Ala2705 2710 2715Gly Leu Ala Ala His Thr Trp Asn Gly Val Ser Leu His Ala Arg2720 2725 2730Gly Ala Thr Val Leu Arg Leu Arg Met Thr Pro Thr Asp Ala Thr2735 2740 2745Ser Thr Ala Val Arg Leu His Ala Thr Asp Glu Thr Gly Ala Pro2750 2755 2760Val Leu Thr Leu Asp Ser Leu Leu Met Arg Pro Val Pro Leu Glu2765 2770 2775
Gly Leu Gly Ala Gly Val Arg Arg Gly Ser Leu Phe Glu Leu Gly2780 2785 2790Trp Val Pro Val Glu Gly Met Pro Ala Ser Val Ala Gly Gly Gly2795 2800 2805Gly Glu Leu Val Ala Trp Glu Cys Pro Gly Gly Gly Val Ala Glu2810 2815 2820Val Thr Ala Ala Ala Leu Gly Val Val Gln Glu Trp Leu Ala Asp2825 2830 2835Glu Arg Glu Gly Asp Ala Arg Leu Val Val Val Thr Arg Gly Ala2840 2845 2850Val Ala Val Asp Ala Gly Glu Pro Val Arg Asp Val Ala Gly Ala2855 2860 2865Ala Val Trp Gly Leu Val Arg Ser Ala Gln Ser Glu His Pro Asp2870 2875 2880Arg Phe Ala Leu Leu Asp Leu Asp Pro Asp Thr Lys Thr Asp Pro2885 2890 2895Gly Ile Asp Thr Asp Gly Asp Thr Asp Val Ser Ala Asp Ala Lys2900 2905 2910Val Gly Thr Gly Asp Gly Leu Asp Asp Ala Ala Val Ala Ser Ala2915 2920 2925Leu Ala Arg Gly Glu Ser Gln Leu Ala Val Arg Asp Gly Val Val2930 2935 2940
Arg Val Ala Arg Leu Gly Gly Leu Val Gly Gly Leu Ser Leu Pro2945 2950 2955Gly Gly Val Gly Trp Arg Leu Asp Gly Gly Gly Ser Gly Leu Leu2960 2965 2970Glu Gly Val Gly Val Val Ala Ser Asp Ala Ala Gly Val Val Leu2975 2980 2985Gly Arg Gly Gln Val Arg Val Ala Val Arg Ala Ala Gly Val Asn2990 2995 3000Phe Arg Asp Val Leu Val Ala Leu Gly Met Val Pro Gly Gln Val3005 3010 3015Gly Val Gly Ser Glu Gly Ala Gly Val Val Val Glu Val Gly Pro3020 3025 3030Gly Val Glu Gly Leu Val Val Gly Asp Arg Val Phe Gly Val Phe3035 3040 3045Gly Asp Ala Phe Ala Pro Val Val Val Ala Gln Glu Val Leu Leu3050 3055 3060Ala Arg Ile Pro Glu Gly Trp Ser Phe Ala Gln Ala Ala Ser Val3065 3070 3075Pro Val Val Phe Ala Thr Ala Tyr Leu Gly Leu Val Asp Leu Ala3080 3085 3090Gly Val Arg Arg Gly Glu Ser Val Leu Val His Ala Ala Ala Gly3095 3100 3105
Gly Val Gly Thr Ala Ala Val Gln Leu Ala Arg His Leu Gly Ala3110 3115 3120Glu Val Tyr Ala Thr Ala Ser Glu Ala Lys Trp Ala Arg Leu Arg3125 3130 3135Ala Ala Gly Val Ala Pro Gln Arg Ile Ala Ser Ser Arg Ser Val3140 3145 3150Glu Phe Glu Ser Arg Phe Arg Arg Ala Ser Gly Gly Arg Gly Val3155 3160 3165Asp Val Val Leu Asn Cys Leu Ala Gly Glu Tyr Thr Asp Ala Ser3170 3175 3180Leu Arg Leu Cys Ser Pro Gln Gly Gly Arg Phe Leu Glu Leu Gly3185 3190 3195Lys Thr Asp Ile Arg Asp Ala Gly Glu Val Ala Ala Arg Phe Pro3200 3205 3210Gly Val Ser Tyr Arg Ala Tyr Asp Leu Met Asp Ala Gly Ala Gln3215 3220 3225Arg Val Gly Glu Ile Leu His Thr Val Val Asp Leu Phe Arg Arg3230 3235 3240Gly Val Leu Glu Pro Leu Pro Val Thr Ala Trp Asp Val Arg Gln3245 3250 3255Ala His Gln Ala Leu Arg Ser Met Arg Ser Gly Leu His Val Gly3260 3265 3270
Lys Asn Val Leu Thr Leu Pro Val Pro Leu Asp Ala Glu Gly Thr3275 3280 3285Val Leu Val Thr Gly Gly Thr Gly Thr Leu Gly Ala Ala Val Ala3290 3295 3300Arg His Leu Ala Ala Gly His Gly Val Arg His Leu Leu Leu Val3305 3310 3315Ser Arg Arg Gly Met Ala Ala Ala Gly Ala Glu Lys Leu Cys Ala3320 3325 3330Glu Leu Gly Gln Ala Gly Val Ser Val Ser Val Ala Gly Cys Asp3335 3340 3345Val Ala Asp Arg Ala Gln Val Ala Ala Leu Leu Glu Gln Val Pro3350 3355 3360Ala Glu His Pro Leu Thr Ala Val Val His Thr Ala Gly Val Leu3365 3370 3375Asp Asp Ala Thr Val Thr Cys Leu Asp Arg Asn Lys Ile Asp Ala3380 3385 3390Val Leu Gly Ala Lys Val Asp Gly Ala Leu His Leu His Glu Leu3395 3400 3405Thr Ala Gly Met Asp Leu Ser Ala Phe Val Leu Phe Ser Ser Ala3410 3415 3420Ala Gly Val Leu Gly Ser Pro Gly Gln Gly Asn Tyr Ala Ala Ala3425 3430 3435
Asn Ala Ala Leu Asp Ala Leu Ala His Gln Arg Arg Ala Ala Gly3440 3445 3450Leu Pro Ala Leu Ser Leu Ala Trp Gly Leu Trp Glu Glu Ala Ser3455 3460 3465Gly Met Thr Gly His Leu Asp Ala Ala Asp Arg His Arg Ile Thr3470 3475 3480Arg Ser Gly Leu His Pro Leu Thr Thr Pro Asp Ala Leu Ala Leu3485 3490 3495Leu Asp Thr Ala Leu Ala Ala Gly Arg Pro Ala Leu Leu Pro Ala3500 3505 3510Asp Leu Arg Pro Thr His Pro Ala Pro Pro Leu Leu Glu His Leu3515 3520 3525Ala Pro Ala Arg Thr Ser His Arg Thr Ala His Thr Ser Thr Ala3530 3535 3540Thr Gly Val Gly Gln Asp Val Ser Leu Thr Asp Arg Leu Ala Thr3545 3550 3555Leu Thr Pro Glu Gln Arg His Asp Thr Leu Leu Ala Leu Ala Arg3560 3565 3570Thr His Ile Ala Ala Val Leu Gly His Pro Ser Pro Asp Thr Ile3575 3580 3585Asp Pro Glu Arg Thr Phe Arg Asp Leu Gly Phe Asp Ser Leu Thr3590 3595 3600
Ala Val Glu Leu Arg Asn Arg Leu Thr Arg Ala Thr Gly Leu Arg3605 3610 3615Leu Pro Ala Thr Leu Ala Phe Asp His Pro Thr Pro Thr Ala Leu3620 3625 3630Thr His His Leu Thr Thr Leu Leu Asn Pro Asn Asp Asn Asp Asn3635 3640 3645Val Gly Pro Val Leu Met Glu Leu Glu Arg Leu Glu Ser Ala Leu3650 3655 3660Ala Ala Leu Asp Arg Asp Asp Ser Ala Cys Glu Arg Val Thr Leu3665 3670 3675Arg Leu Gln Ser Leu Met Leu Arg Trp Ser Gly Ser Glu Arg Gln3680 3685 3690Ser Ala Glu Asn Thr Asp Asp Ser Ser Arg Phe Ala Ser Ala Thr3695 3700 3705Ala Glu Glu Leu Leu Glu Phe Ile Asp Arg Asp Leu Gly Leu Ser3710 3715 3720<210>7<211>6043<212>PRT<213>细菌<400>7Val Ala Asn Asp Glu Lys Val Leu Glu Tyr Leu Lys Arg Val Thr Ala1 5 10 15Asp Leu Asp Arg Thr Arg Arg Arg Leu Tyr Glu Val Val Glu Arg Glu
20 25 30Gln Glu Pro Ile Ala Ile Val Gly Met Ala Cys Arg Tyr Pro Gly Gly35 40 45Ala Gly Ser Pro Ala Gly Leu Trp Asp Leu Val Ser Ser Gly Thr Asp50 55 60Ala Ile Gly Glu Phe Pro Thr Asp Arg Gly Trp Asp Leu Glu Arg Leu65 70 75 80Tyr Asp Pro Asp Pro Asp His Pro Gly Thr Thr Tyr Thr Arg His Gly85 90 95Gly Phe Leu Asp Gly Val Gly Glu Phe Asp Ala Glu Phe Phe Gly Val100 105 110Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Leu Leu115 120 125Glu Thr Ala Trp Glu Ala Ile Glu His Ala Gly Ile Val Pro Glu Ser130 135 140Leu Arg Gly Thr Ser Thr Gly Val Phe Ala Gly Ile Asn Pro Gln Asp145 150 155 160Tyr Thr Ile Ser Gln Tyr Gly Arg Asp Ser Glu Ile Glu Gly Tyr Leu165 170 175Leu Thr Gly Ala Ala Ala Ser Ile Ala Ser Gly Arg Ile Ser Tyr Thr180 185 190Leu Gly Leu Glu Gly Pro Ala Val Thr Ile Asp Thr Ala Cys Ser Ser
195 200 205Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala Leu Arg Ala Gly Glu210 215 220Cys Thr Met Ala Leu Ala Gly Gly Ala Ser Val Leu Ser Thr Pro Leu225 230 235 240Ile Phe Val Glu Phe Ala Arg His His Gly Leu Ser Val Asp Gly Arg245 250 255Cys Lys Ala Phe Ser Ala Ser Ala Asp Gly Thr Gly Trp Gly Glu Gly260 265 270Ala Gly Leu Leu Leu Leu Glu Arg Leu Ser Asp Ala Lys Arg Asn Gly275 280 285Arg Arg Ile Leu Ala Leu Val Arg Gly Ser Ala Val Asn Gln Asp Gly290 295 300Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Cys Arg Val305 310 315 320Ile Arg Arg Ala Leu Ala Asn Ala His Leu Ala Pro Ala Asp Ile Asp325 330 335Ala Val Glu Ala His Gly Thr Gly Thr Thr Leu Gly Asp Pro Ile Glu340 345 350Ala Gln Ala Leu Gln Glu Ala Tyr Gly Ala Asp Arg Pro Asp Asp Arg355 360 365Pro Leu Trp Val Gly Thr Leu Lys Ser Asn Ile Gly His Ser Ile Ala
370 375 380Ala Ala Gly Val Gly Gly Val Ile Lys Met Val Met Ala Leu Arg His385 390 395 400Glu Ser Leu Pro Arg Thr Leu His Val Asp Glu Pro Ser Pro Gln Val405 410 415Asp Trp Ser Ser Gly Ala Val Ser Leu Leu Thr Glu Ala Arg Pro Trp420 425 430Pro Arg Asp Glu Asp Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gly435 440 445Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Glu Ala Pro Ala Pro450 455 460Ala Glu Val Gln Ala Val Glu Thr Ala Pro Val Val Arg Val Asp Gly465 470 475 480Gly Glu Arg Ser Ala Pro Ala Asp Val Pro Leu Val Trp Val Val Ser485 490 495Gly Lys Ser Gln Ala Ala Leu Arg Ala Gln Ala Ala Ala Leu His Ala500 505 510His Val Leu Asp His Pro Glu Gln Asp Ala Ala Asp Ile Gly Tyr Ser515 520 525Leu Ala Thr Thr Arg Ala Leu Phe Asp His Arg Ala Thr Leu Ile Ala530 535 540Pro Asp Arg Asp Thr Leu Leu Asp Ala Leu Thr Ala Leu Ala Asp Gly
545 550 555 560Arg Thr His Pro His Leu Val Pro Ala Pro Pro Thr Glu Pro Gly His565 570 575Ala His Lys Ile Ala Phe Leu Cys Ser Gly Gln Gly Thr Gln Arg Pro580 585 590Gly Met Ala Thr Gly Leu Tyr His Thr Tyr Pro Ala Phe Ala Ala Ala595 600 605Leu Asp Glu Thr Cys Ala His Phe Asp Pro His Leu Asp His Pro Leu610 615 620Arg Asp Leu Leu Leu Asn His Asp Pro Thr Gly Leu Leu Thr His Thr625 630 635 640Leu Tyr Ala Gln Pro Ala Leu Phe Thr Leu Gln Lys Ala Leu His His645 650 655Leu Ile Thr Glu Thr Tyr Gly Ile Thr Pro His Tyr Leu Ala Gly His660 665 670Ser Leu Gly Glu Ile Thr Ala Ala His Leu Ala Gly Ile Leu Thr Leu675 680 685Pro Asp Ala Thr His Leu Ile Thr Thr Arg Ala Arg Leu Met Gln Thr690 695 700Met Pro Pro Gly Thr Met Thr Thr Leu His Thr Thr Pro Glu His Ile705 710 715 720Gln Pro Leu Leu Asp Gln His Pro Gly Lys Ala Thr Ile Ala Ala Val
725 730735Asn Ser Pro His Ser Leu Val Ile Ser Gly Asp Pro Asp Thr Ile His740 745 750His Ile Thr Thr Thr Cys His Thr Gln Gly Ile Thr Thr Lys Pro Leu755 760 765Thr Thr Asn His Ala Phe His Ser Pro His Thr Asp Thr Ile Leu Glu770 775 780Gln Leu Asp Thr Thr Thr His Thr Leu Thr Tyr His Pro Pro His Thr785 790 795 800Pro Leu Ile Thr Ser Thr Pro Gly Asp Pro Leu Thr Pro His Tyr Trp805 810 815Thr His Gln Thr Arg Gln Pro Val His Trp Thr Asp Thr Ile His Thr820 825 830Leu His Thr Asn Gly Val Thr Thr Tyr Ile Glu Leu Gly Pro Asp His835 840 845Thr Leu Thr Thr Leu Thr His His Asn Leu Pro His His Gln Pro Thr850 855 860Ala Ile Thr Leu Thr His Pro His His Asn Pro Thr His His Leu Leu865 870 875 880Thr Ala Leu Ala His Thr Pro Thr Thr Trp His Thr His His His Thr885 890 895His Thr Asn Pro His Pro His Thr Ile Pro Asp Leu Pro Thr Tyr Pro
900 905 910Phe Gln Arg Arg His Tyr Trp Leu Gln Ala Pro Thr Thr Ser Thr Asp915 920 925Gln Pro Val Ala Pro Thr Asn Asp Asp Ala Pro Ala Pro Arg Ala Thr930 935 940Ser Leu Arg Asp Thr Leu Ala Gly Arg Ser Pro Gln Glu Arg Glu Glu945 950 955 960Val Leu Leu Asp Leu Val Leu Thr Gln Val Ala Ala Val Leu Gly His965 970 975Thr Ala Pro Glu Val Val Asp Pro Gln Arg Ala Phe Lys Asp Leu Gly980 985 990Phe Asp Ser Leu Ala Ala Ile Lys Leu Arg Asn Arg Leu Ala Ala Ala995 1000 1005Thr Gly Leu Glu Leu Pro Thr Thr Leu Val Phe Asp His Pro Thr1010 1015 1020Pro Val Ala Leu Arg Gln Tyr Phe Gln Ser Gln Ile Leu Gly Ala1025 1030 1035Glu Ala Asp Ala Pro Asn Arg Leu Pro Leu Arg Ala Ala Thr Thr1040 1045 1050Asp Glu Pro Ile Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly1055 1060 1065Gly Val Arg Thr Ala Asp Asp Leu Trp Gln Leu Leu Ser Asp Glu
1070 1075 1080His Asp Ala Val Gly Gly Phe Pro Thr Asn Arg Gly Trp Asp Val1085 1090 1095Ala Asn Leu Tyr Asp Pro Asp Pro Asp Arg His Gly Thr Thr Tyr1100 1105 1110Thr Gln Gln Gly Gly Phe Leu Tyr Glu Ala Gly Glu Phe Asp Ala1115 1120 1125Glu Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro1130 1135 1140Gln Gln Arg Leu Leu Leu Glu Thr Ala Trp Glu Ala Ile Glu His1145 1150 1155Ala Gly Ile Asn Pro Asp Ala Leu Arg Asn Thr Ser Thr Gly Val1160 1165 1170Phe Ala Gly Val Ile Tyr His Asp Tyr Ala Ser Arg Phe Leu Thr1175 1180 1185Ala Pro Ala Gly Tyr Glu Gly Tyr Leu Gly His Gly Ser Ala Gly1190 1195 1200Ser Ile Ala Ser Gly Arg Val Ala Tyr Val Leu Gly Leu Glu Gly1205 1210 1215Pro Ala Val Thr Val Asp Thr Ala Cys Ser Ser Ser Leu Val Ala1220 1225 1230Leu His Leu Ala Cys Gln Ala Leu Arg Ser Gly Glu Cys Thr Met
1235 1240 1245Ala Leu Ala Gly Gly Ala Thr Val Met Ser Thr Pro Gln Ala Phe1250 1255 1260Val Glu Phe Ser Arg Gln Arg Gly Leu Ala Ala Asp Gly Arg Cys1265 1270 1275Lys Ala Phe Ser Ala Ala Ala Asp Gly Thr Gly Trp Gly Glu Gly1280 1285 1290Ala Gly Leu Leu Leu Leu Glu Arg Leu Ser Glu Ala Glu Arg Asn1295 1300 1305Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln1310 1315 1320Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln1325 1330 1335Gln Arg Val Ile Arg Gln Ala Leu Ala Asn Ser Gly Leu Thr Gly1340 1345 1350Ala Asp Val Asp Ala Val Glu Ala His Gly Thr Gly Thr Lys Leu1355 1360 1365Gly Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln1370 1375 1380Glu His His Pro Asp Gln Pro Leu Trp Leu Gly Ser Leu Lys Ser1385 1390 1395Asn Ile Gly His Ala Gln Ala Ala Ala Gly Val Gly Ser Ile Ile
1400 1405 1410Lys Met Ile Met Ala Met Arg Asn Glu Ser Leu Pro Arg Thr Leu1415 1420 1425His Val Asp Glu Pro Ser Pro His Val Asp Trp Ser Ser Gly Ala1430 1435 1440Val Ser Leu Leu Thr Glu Pro Arg Pro Trp Pro Arg Arg Glu Asp1445 1450 1455Arg Pro Arg Arg Ala Gly Ile Ser Ser Phe Gly Val Ser Gly Thr1460 1465 1470Asn Ala His Val Ile Val Glu Glu Pro Pro Ala Arg Ala Glu Val1475 1480 1485Glu Ala Val Glu Ala Ala Pro Ala Gly Val Glu Thr Ala Ala Ala1490 1495 1500Ala Ala Val Val Val Glu Thr Asp Gly Ala Gly Arg Val Ser Ser1505 1510 1515Asp Val Pro Leu Val Trp Val Val Ser Gly Lys Ser Gln Ala Ala1520 1525 1530Leu Arg Ala Gln Ala Ala Ala Leu His Ala His Val Leu Asp His1535 1540 1545Pro Glu Gln Asp Ala Ala Asp Ile Gly Tyr Ser Leu Ala Thr Thr1550 1555 1560Arg Ala Leu Phe Asp His Arg Ala Thr Leu Ile Ala Pro Asp Arg
1565 1570 1575Asp Thr Leu Leu Asp Ala Leu Thr Ala Leu Ala Asp Gly Arg Thr1580 1585 1590His Pro His Leu Ile Pro Thr Pro Pro Thr Glu Pro Gly His Thr1595 1600 1605His Lys Ile Ala Phe Leu Cys Ser Gly Gln Gly Thr Gln Arg Pro1610 1615 1620Gly Met Ala Thr Gly Leu Tyr His Thr Tyr Pro Ala Phe Ala Ala1625 1630 1635Ala Leu Asp Glu Thr Cys Ala His Phe Asp Pro His Leu Asp His1640 1645 1650Pro Leu Arg Asp Leu Leu Leu Asn His Asp Pro Thr Asp Leu Leu1655 1660 1665Thr His Thr Leu Tyr Ala Gln Pro Ala Leu Phe Thr Leu Gln Lys1670 1675 1680Ala Leu His His Leu Ile Thr Glu Thr Tyr Gly Ile Thr Pro His1685 1690 1695Tyr Leu Ala Gly His Ser Leu Gly Glu Ile Thr Ala Ala His Leu1700 1705 1710Ala Gly Ile Leu Thr Leu Pro Asp Ala Thr His Leu Ile Thr Thr1715 1720 1725Arg Ala Arg Leu Met Gln Thr Met Pro Pro Gly Thr Met Thr Thr
1730 1735 1740Leu His Thr Thr Pro Glu His Ile Gln Pro Leu Leu Asp Gln His1745 1750 1755Pro Gly Lys Ala Thr Ile Ala Ala Val Asn Ser Pro His Ser Leu1760 1765 1770Val Ile Ser Gly Asp Pro Asp Thr Ile His His Ile Thr Thr Thr1775 1780 1785Cys His Asn Gln Gly Ile Thr Thr Lys Pro Leu Thr Thr Asn His1790 1795 1800Ala Phe His Ser Pro His Thr Asn Thr Ile Leu Glu Gln Leu Asp1805 1810 1815Thr Thr Thr His Thr Leu Thr Tyr His Pro Pro His Thr Pro Leu1820 1825 1830Ile Thr Ser Thr Pro Gly Asn Pro Leu Thr Pro His Tyr Trp Thr1835 1840 1845His Gln Thr Arg Gln Pro Val His Trp Ala Asp Thr Ile His Thr1850 1855 1860Leu His Thr Asn Gly Val Thr Thr Tyr Ile Gly Leu Gly Pro Asp1865 1870 1875His Thr Leu Ser Thr Leu Thr His His Asn Leu Pro Gln His Gln1880 1885 1890Pro Thr Ala Ile Thr Leu Thr His Pro His His Asn Pro Thr His
1895 1900 1905His Leu Leu Thr Ala Leu Ala His Thr Pro Thr Thr Trp His Thr1910 1915 1920His His His Thr His Thr Asn Pro His Pro His Thr Ile Pro Asp1925 1930 1935Leu Pro Thr Tyr Pro Phe Gln Arg Arg His Tyr Trp Leu Glu Val1940 1945 1950Pro Lys Pro Thr Ala Glu Ala Ser Ala Ser Ala Ser Gly Pro Gly1955 1960 1965Arg Asn Arg Ala Ala Lys Leu Ser Ala Leu Glu Ala Glu Phe Trp1970 1975 1980Gln Ala Val Glu Glu Thr Asp Thr Asp Thr Leu Ala His Thr Leu1985 1990 1995Asp Leu Asp Thr Gln Thr Leu Glu Pro Val Leu Pro Ala Leu Ala2000 2005 2010Thr Trp His Gln Gln Gln Arg Asp His Ala Arg Ile Asn Thr Trp2015 2020 2025Thr Tyr Gln Glu Thr Trp Lys Pro Leu His Leu Pro Thr Thr Arg2030 2035 2040Pro Thr Thr Pro Thr Ser Trp Leu Ile Ala Ile Pro Glu Thr His2045 2050 2055Arg Asn His Pro His Thr Thr Asn Leu Leu Thr Asn Leu Pro His
2060 2065 2070His Asn Ile Thr Pro Ile Pro Leu Thr Ile Asn His Thr Thr Asp2075 2080 2085Leu His His Ala Tyr His His Ala His His His Thr Thr Pro Pro2090 2095 2100Ile Thr Ala Val Leu Ser Leu Leu Ala Leu Asp Glu Thr Pro His2105 2110 2115Pro His His Pro His Thr Pro Thr Gly Thr Leu Leu Asn Leu Thr2120 2125 2130Leu Thr Gln Thr His Thr Gln Thr His Pro Pro Thr Pro Leu Trp2135 2140 2145Tyr Leu Thr Thr Gln Ala Thr Thr Thr His Pro Asn Asp Pro Leu2150 2155 2160Thr His Pro Thr Gln Ala Gln Thr Ile Gly Leu Ala Arg Thr Thr2165 2170 2175His Leu Glu His Pro His His Thr Gly Gly His Ile Asp Leu Pro2180 2185 2190Thr Thr Pro His Pro Asn Thr Leu Thr Gln Leu Ile Thr Ala Leu2195 2200 2205Thr His Pro His His Gln His Asn Leu Thr Ile Arg Thr His Thr2210 2215 2220Thr His Thr Arg Arg Leu Thr Pro Thr Thr Leu Gln Pro Thr Thr
2225 2230 2235Pro Thr Pro Pro Thr Asn Pro His Gly Thr Thr Leu Ile Thr Gly2240 2245 2250Gly Thr Gly Ala Leu Ala Thr Thr Leu Ala His His Leu Ala Thr2255 2260 2265Thr Gly Thr Gln His Leu Leu Leu Thr Ser Arg Arg Gly Pro His2270 2275 2280Thr Pro Gly Ala Arg Gln Leu His Thr Gln Leu Thr Gln Leu Gly2285 2290 2295Thr Asn Thr Thr Ile Thr Ala Cys Asp Leu Ser Asp Pro Asp Gln2300 2305 2310Leu Thr His Leu Leu Thr His Ile Pro Pro Glu His Pro Leu Thr2315 2320 2325Thr Val Ile His Thr Ala Gly Ile Leu Asp Asp Ala Thr Leu Thr2330 2335 2340Asn Leu Thr Pro Thr Gln Leu Asp Asn Val Leu Arg Ala Lys Ala2345 2350 2355His Thr Ala His Leu Leu His His Ala Thr Leu His Thr Pro Leu2360 2365 2370Asp His Phe Val Leu Tyr Ser Ser Ala Ala Ala Thr Leu Gly Ala2375 2380 2385Pro Gly Gln Ala Asn Tyr Ala Ala Ala Asn Ala Tyr Leu Asp Ala
2390 2395 2400Leu Ala His His Arg His Thr His Asn Leu Pro Ala Thr Thr Ile2405 2410 2415Ala Trp Gly Thr Trp Gln Gly Asn Gly Leu Ala Ser Gly Asp Ile2420 2425 2430Gly Glu His Leu Arg Arg Arg Gly Met Ile Pro Leu Asp Pro Glu2435 2440 2445Ser Ala Val Gly Ala Phe Asp Arg Ala Val Ala Ser Asp Arg Pro2450 2455 2460Ser Val Phe Val Ala Asp Ile Asp Trp Pro Thr Phe Gly Arg Asn2465 2470 2475Thr Ser Ser Gly Leu Arg Ala Leu Phe Glu Asp Ile Pro Glu Ala2480 2485 2490Thr Gln Pro Glu Pro Thr Ala Arg Ser Ala Asp Gln Pro Asn Gly2495 2500 2505His Gly Ser Leu Gln Glu Leu Leu Ala Arg Gln Ser Pro Ala Glu2510 2515 2520Gln Ala Glu Thr Leu Leu Ala Leu Val Arg Thr His Ser Ala Thr2525 2530 2535Val Leu Gly Arg Asp Gly Ala Asp Ala Val Ala Ala Glu Arg Pro2540 2545 2550Phe Arg Asp Leu Gly Phe Asp Ser Leu Ser Ala Val Glu Leu Arg
2555 2560 2565Asn His Leu Thr Ala Asp Thr Glu Leu Ala Leu Pro Thr Thr Leu2570 2575 2580Val Phe Asp His Pro Thr Pro Val Lys Leu Ala Glu Phe Leu Arg2585 2590 2595Thr Glu Leu Leu Gly Thr Ala Pro Ala Thr Thr Ala Ala Val Pro2600 2605 2610Ala Leu Gln Ser His Thr Asp Glu Pro Ile Ala Ile Ile Gly Met2615 2620 2625Ala Cys Arg Phe Pro Gly Ala Val Thr Thr Pro Glu His Leu Trp2630 2635 2640Asn Leu Ile Ala Thr Glu Gln Asp Ala Ile Gly Glu Phe Pro Thr2645 2650 2655Asp Arg Gly Trp Asp Leu Asp Asn Leu Tyr His Pro Asp Pro Asp2660 2665 2670His Pro Gly Thr Thr Tyr Thr Arg His Gly Gly Phe Leu Tyr Asp2675 2680 2685Ala Gly Asp Phe Asp Ala Glu Phe Phe Gly Ile Asn Pro Arg Glu2690 2695 2700Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Thr Ala2705 2710 2715Trp Glu Ala Ile Glu His Ala Gly Ile Leu Pro Asp Ala Leu His
2720 2725 2730Gly Thr Pro Thr Gly Val Phe Thr Gly Val Asn Ala Gln Asp Tyr2735 2740 2745Ala Ala His Thr His Ala Ser Pro His Thr Thr Glu Gly Tyr Thr2750 2755 2760Leu Thr Gly Thr Ala Gly Ser Ile Ala Ser Gly Arg Ile Ala Tyr2765 2770 2775Thr Leu Gly Leu Glu Gly Pro Ala Val Thr Ile Asp Thr Ala Cys2780 2785 2790Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ala Leu Arg2795 2800 2805Ala Gly Glu Cys Thr Thr Ala Leu Ala Ser Gly Ile Thr Val Met2810 2815 2820Thr Ser Pro Val Thr Phe Thr Glu Phe Ser Arg Gln Arg Gly Leu2825 2830 2835Ala Pro Asp Gly His Cys Lys Ala Phe Ser Ala Ser Ala Asp Gly2840 2845 2850Thr Gly Trp Ser Glu Gly Val Gly Thr Ile Leu Val Glu Arg Leu2855 2860 2865Ser Asp Ala Glu Arg Asn Gly His Arg Ile Leu Ala Val Val Arg2870 2875 2880Gly Ser Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala
2885 2890 2895Pro Asn Gly Pro Ser Gln Gln Arg Val Ile Arg Gln Ala Leu Ala2900 2905 2910Asn Ser Gly Leu Thr Gly Ala Asp Val Asp Ala Val Glu Ala His2915 2920 2925Gly Thr Gly Thr Lys Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu2930 2935 2940Leu Ala Thr Tyr Gly Gln Gly Arg Ala Gln Glu Gln Pro Leu Trp2945 2950 2955Leu Gly Ser Val Lys Ser Asn Leu Gly His Thr Gln Ala Ala Ala2960 2965 2970Gly Met Ala Gly Leu Ile Lys Met Val Met Ala Leu Arg His Glu2975 2980 2985Ser Leu Pro Arg Thr Leu His Val Asp Glu Pro Ser Pro Gln Val2990 2995 3000Asp Trp Ser Ser Gly Ala Val Ser Leu Leu Thr Glu Ala Arg Pro3005 3010 3015Trp Pro Arg Arg Glu Asp Arg Pro Arg Arg Ala Gly Ile Ser Ser3020 3025 3030Phe Gly Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Glu Ala3035 3040 3045Pro Ala Pro Ala Glu Ala Val Glu Thr Glu Gln Gly Val Val Pro
3050 3055 3060Gln Gly Asp Gln Glu Cys Ser Ala Pro Val Gly Val Pro Leu Val3065 3070 3075Trp Val Val Ser Gly Lys Ser Gln Ala Ala Leu Arg Ala Gln Ala3080 3085 3090Ala Ala Leu His Ala His Val Leu Asp His Pro Glu Gln Asp Ala3095 3100 3105Ala Asp Ile Gly Tyr Ser Leu Ala Thr Thr Arg Ala Leu Phe Asp3110 3115 3120His Arg Ala Thr Leu Ile Ala Pro Asp Arg Asp Thr Leu Leu Asp3125 3130 3135Ala Leu Thr Ala Leu Ala Asp Gly Arg Thr His Pro His Leu Ile3140 3145 3150Pro Thr Pro Pro Thr Glu Pro Gly His Thr His Lys Ile Ala Phe3155 3160 3165Leu Cys Ser Gly Gln Gly Thr Gln Arg Pro Gly Met Ala Thr Gly3170 3175 3180Leu Tyr His Thr Tyr Pro Ala Phe Ala Ala Ala Leu Asp Glu Thr3185 3190 3195Cys Ala His Phe Asp Pro His Leu Asp His Pro Leu Arg Asp Leu3200 3205 3210Leu Leu Asn His Asp Pro Thr Asp Leu Leu Thr His Thr Leu Tyr
3215 3220 3225Ala Gln Pro Ala Leu Phe Thr Leu Gln Lys Ala Leu His His Leu3230 3235 3240Ile Thr Glu Thr Tyr Gly Ile Thr Pro His Tyr Leu Ala Gly His3245 3250 3255Ser Leu Gly Glu Ile Thr Ala Ala His Leu Ala Gly Ile Leu Thr3260 3265 3270Leu Pro Asp Ala Thr His Leu Ile Thr Thr Arg Ala Arg Leu Met3275 3280 3285Gln Thr Met Pro Pro Gly Thr Met Thr Thr Leu His Thr Thr Pro3290 3295 3300Glu His Ile Gln Pro Leu Leu Asp Gln His Pro Gly Lys Ala Thr3305 3310 3315Ile Ala Ala Val Asn Ser Pro His Ser Leu Val Ile Ser Gly Asp3320 3325 3330Pro Asp Thr Ile His His Ile Thr Thr Thr Cys His Thr Gln Gly3335 3340 3345Ile Thr Thr Lys Pro Leu Thr Thr Asn His Ala Phe His Ser Pro3350 3355 3360His Thr Asp Thr Ile Leu Glu Gln Leu Asp Thr Thr Thr His Thr3365 3370 3375Leu Thr Tyr His Gln Pro His Thr Pro Leu Ile Thr Ser Thr Pro
3380 3385 3390Gly Asp Pro Leu Thr Pro His Tyr Trp Thr His Gln Thr Arg Gln3395 3400 3405Pro Val His Trp Ala Asp Thr Ile His Thr Leu His Thr Asn Gly3410 3415 3420Val Thr Thr Tyr Ile Gly Leu Gly Pro Asp His Thr Leu Ser Thr3425 3430 3435Leu Thr His His Asn Leu Pro Gln His Gln Pro Thr Ala Ile Thr3440 3445 3450Leu Thr His Pro His His Asn Pro Thr His His Leu Leu Thr Ala3455 3460 3465Leu Ala His Thr Pro Thr Thr Trp His Thr His His His Thr His3470 3475 3480Thr Asn Pro His Pro His Thr Ile Pro Asp Leu Pro Thr Tyr Pro3485 3490 3495Phe Gln Arg Arg His Tyr Trp Leu Glu Val Pro Lys Pro Thr Ala3500 3505 3510Glu Ala Ser Ala Ser Ala Ser Gly Pro Gly Arg Asn Arg Ala Ala3515 3520 3525Lys Leu Ser Ala Leu Glu Ala Glu Phe Trp Gln Ala Val Glu Glu3530 3535 3540Thr Asp Thr Asp Thr Leu Ala His Thr Leu Asp Leu Asp Thr Gln
3545 3550 3555Thr Leu Glu Pro Val Leu Pro Ala Leu Ala Thr Trp His Gln Gln3560 3565 3570Gln Arg Asp His Ala Arg Ile Asn Thr Trp Thr Tyr Gln Glu Thr3575 3580 3585Trp Lys Pro Leu His Leu Pro Thr Thr Arg Pro Thr Thr Pro Thr3590 3595 3600Ser Trp Leu Ile Ala Ile Pro Glu Thr His Arg Asn His Pro His3605 3610 3615Thr Thr Asn Leu Leu Thr Asn Leu Pro His His Asn Ile Thr Pro3620 3625 3630Ile Pro Leu Thr Ile Asn His Thr Thr Asp Leu His His Ala Tyr3635 3640 3645His His Ala His His His Thr Thr Pro Pro Ile Thr Ala Val Leu3650 3655 3660Ser Leu Leu Ala Leu Asp Glu Thr Pro His Pro His His Pro His3665 3670 3675Thr Pro Thr Gly Thr Leu Leu Asn Leu Thr Leu Thr Gln Thr His3680 3685 3690Thr Gln Thr His Pro Pro Thr Pro Leu Trp Tyr Leu Thr Thr Gln3695 3700 3705Ala Thr Thr Thr His Pro Asn Asp Pro Leu Thr His Pro Thr Gln
3710 3715 3720Ala Gln Thr Ile Gly Leu Ala Arg Thr Thr His Leu Glu His Pro3725 3730 3735His His Thr Gly Gly His Ile Asp Leu Pro Thr Thr Pro His Pro3740 3745 3750Asn Thr Leu Thr Gln Leu Ile Thr Ala Leu Thr His Pro His His3755 3760 3765Gln His Asn Leu Thr Ile Arg Thr His Thr Thr His Thr Arg Arg3770 3775 3780Leu Thr Pro Thr Thr Leu Gln Pro Thr Thr Pro Thr Pro Pro Thr3785 3790 3795Asn Pro His Gly Thr Thr Leu Ile Thr Gly Gly Thr Gly Ala Leu3800 3805 3810Ala Thr Thr Leu Ala His His Leu Ala Thr Thr Gly Thr Gln His3815 3820 3825Leu Leu Leu Thr Ser Arg Arg Gly Pro His Thr Pro Gly Ala Arg3830 3835 3840Gln Leu His Thr Gln Leu Thr Gln Leu Gly Thr Asn Thr Thr Ile3845 3850 3855Thr Ala Cys Asp Leu Ser Asp Pro Asp Gln Leu Thr His Ile Leu3860 3865 3870Thr His Ile Pro Pro Glu His Pro Leu Thr Thr Val Ile His Thr
3875 3880 3885Ala Gly Val Asn His Tyr Ala Pro Val Ala Ala Thr Asp Pro Ser3890 3895 3900Thr Phe Ala Ser Val Leu Ala Ala Lys Ala Ala Gly Ala Ala His3905 3910 3915Leu His Glu Leu Leu Leu Glu Leu Asp Thr Val Glu Gln Phe Ile3920 3925 3930Leu Phe Ser Ser Gly Ser Gly Ala Trp Gly Ser Gly Asn Gln Cys3935 3940 3945Ala Tyr Ala Ala Ala Asn Ala Tyr Leu Asp Ala Leu Ala Ala His3950 3955 3960Arg Gln Ala Arg Gly Leu Pro Gly Met Ser Leu Ala Trp Gly Pro3965 3970 3975Trp Asp Gly Asp Gly Met Ser Ala Gly Glu Asp Ala Gln Arg Tyr3980 3985 3990Leu Arg Glu Arg Gly Val Leu Pro Met Asp Pro Arg Leu Ala Val3995 4000 4005Ala Ala Phe Asp Glu Ala Val Arg Ala Arg Pro Asn Ser Asn Leu4010 4015 4020Val Val Ala Asp Ile Asp Trp Glu Arg Phe Val Pro Thr Phe Thr4025 4030 4035Ala Arg Gly His Asn Pro Leu Ile Glu Asp Ile Pro Glu Val Arg
4040 4045 4050Arg Leu Ala Ala Glu Ala Glu Ala Ala Gln Thr Thr Thr Ala Ala4055 4060 4065Thr Asp Ala Pro Ala Leu Leu Asn Arg Leu Ser Gly Leu Ser Ala4070 4075 4080Thr Gln Gln Lys Gln His Leu Leu Arg Leu Val Arg Ser His Met4085 4090 4095Gly Glu Val Leu Gly Arg Glu Asp Val Asp Thr Leu Asp Glu Arg4100 4105 4110His Thr Phe Arg Asp Leu Gly Phe Asp Ser Leu Thr Ser Ala Arg4115 4120 4125Phe Ser Gln Arg Leu Ala Lys Asp Thr Gly Leu His Leu Pro Ala4130 4135 4140Thr Leu Val Phe Asp His Pro Thr Pro Ala Asp Cys Val Ala His4145 4150 4155Leu Arg Asp Gln Leu Leu Gly Glu Thr Asp Asp Met Thr Pro Arg4160 4165 4170Lys Arg Asp His Leu Gly Glu Asp Arg Arg Ala Ala Thr Ala Asp4175 4180 4185Asp Pro Ile Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly Gly4190 4195 4200Val Arg Ser Ala Asp Asp Leu Trp Asp Leu Leu Ser Ser Gly Thr
4205 4210 4215Asp Ala Ile Ser Gly Phe Pro Thr Asp Arg Gly Trp Asp Ile Glu4220 4225 4230Ser Leu Tyr Asp Pro Asp Pro Asp Arg Ser Gly Thr Thr Tyr Thr4235 4240 4245Arg His Gly Gly Phe Leu Tyr Asp Ala Gly Gln Phe Asp Ala Glu4250 4255 4260Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln4265 4270 4275Gln Arg Leu Leu Leu Glu Thr Ala Trp Glu Ala Val Glu His Ala4280 4285 4290Gly Ile Asn Pro Gln Thr Leu His Gly Thr Pro Thr Gly Val Phe4295 4300 4305Thr Gly Val Asn Ala Gln Asp Tyr Ala Ala His Leu Arg Gln Ala4310 4315 4320Ser Gly Asn Val Glu Gly Tyr Ala Leu Thr Gly Ser Ser Gly Ser4325 4330 4335Val Val Ser Gly Arg Val Ala Tyr Thr Phe Gly Phe Glu Gly Pro4340 4345 4350Ala Val Ser Val Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu4355 4360 4365His Leu Ala Gly Gln Ala Leu Arg Ser Gly Glu Cys Thr Met Ala
4370 4375 4380Leu Ala Gly Gly Val Met Val Met Ser Ser Pro Glu Thr Phe Val4385 4390 4395Glu Phe Ser Arg Gln Arg Gly Leu Ser Val Asp Gly Arg Cys Lys4400 4405 4410Ser Phe Ala Ala Ala Ala Asp Gly Thr Gly Trp Gly Glu Gly Val4415 4420 4425Gly Met Leu Leu Val Glu Arg Leu Ser Asp Ala Glu Arg Asn Gly4430 4435 4440His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp4445 4450 4455Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Gln4460 4465 4470Arg Val Ile Arg Gln Ala Leu Ala Asn Ser Gly Leu Thr Gly Ala4475 4480 4485Asp Val Asp Ala Val Glu Ala His Gly Thr Gly Thr Lys Leu Gly4490 4495 4500Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Glu4505 4510 4515His His Pro Asp Gln Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn4520 4525 4530Ile Gly His Ala Gln Ala Ala Ala Gly Val Gly Gly Ile Ile Lys
4535 4540 4545Met Val Met Ala Leu Arg His Glu Thr Leu Pro Arg Thr Leu His4550 4555 4560Ile Asp Glu Pro Thr Pro Gln Val Asp Trp Ser Ser Gly Ala Val4565 4570 4575Ser Leu Leu Thr Glu Pro Arg Pro Trp Pro Arg Gln Gly Asp Arg4580 4585 4590Pro Arg Arg Ala Gly Ile Ser Ser Phe Gly Val Ser Gly Thr Asn4595 4600 4605Ala His Val Ile Leu Glu Glu Ala Pro Ala Gln Pro Ala Gly Asp4610 4615 4620Pro Ala Pro Glu Asp Gly Ala Pro Val Pro Trp Ala Met Ser Ala4625 4630 4635Arg Ser Asn Ala Ala Leu Arg Ala Gln Ala Ala Leu Leu Arg Asp4640 4645 4650Phe Leu Gln Gly Pro Gly Thr Asp Thr Ala Leu Arg Ala Val Gly4655 4660 4665Ala Glu Leu Ala His Gly Arg Ala Val Leu Glu His Arg Ala Val4670 4675 4680Ile Val Ala Arg Glu Arg Thr Glu Phe Glu Asp Ala Leu Glu Ala4685 4690 4695Leu Ala Ser Gly Glu Pro His Pro Ala Leu Ile Glu Asp Thr Thr
4700 4705 4710Gly Ser Gln Thr Asn Ser His Ser Gly Gly Gly Val Val Phe Val4715 4720 4725Phe Pro Gly Gln Gly Gly Gln Trp Ala Gly Met Gly Leu Asp Leu4730 4735 4740Leu Arg Asp Ser Gln Val Phe Ala Asp His Val Gly Ala Cys Glu4745 4750 4755Arg Ala Leu Ala Pro Trp Val Glu Trp Ser Leu Thr Glu Met Leu4760 4765 4770His Arg Asp Ala Glu Asp Pro Val Trp Glu Arg Ala Asp Val Val4775 4780 4785Gln Pro Val Leu Phe Ser Val Met Val Ser Leu Ala Ala Leu Trp4790 4795 4800Arg Ser Tyr Gly Ile Glu Pro Glu Ala Val Val Gly His Ser Gln4805 4810 4815Gly Glu Ile Ala Ala Ala His Val Cys Gly Ala Leu Thr Leu Glu4820 4825 4830Asp Ala Ala Lys Ile Val Ala Leu Arg Ser Arg Ala Leu Ala Ala4835 4840 4845Leu Arg Gly His Gly Gly Met Ala Ser Leu Ala Leu Thr Gly Thr4850 4855 4860Glu Ala Glu Asp Leu Ile Thr Thr His Trp Pro Gly Arg Leu Trp
4865 4870 4875Thr Ala Ala Phe Asn Gly Pro Arg Ala Thr Thr Val Ser Gly Asp4880 4885 4890Thr Asp Ala Leu Asp Glu Leu Leu Thr His Cys Thr Glu Thr Gly4895 4900 4905Val Arg Ala Arg Arg Ile Pro Val Asp Tyr Ala Ser His Cys Pro4910 4915 4920His Thr Glu Thr Ile Glu His Asp Leu Leu His Met Leu His Gly4925 4930 4935Ile Thr Pro Gln Pro Gly Ser Ile Pro Phe Tyr Ser Thr Val Glu4940 4945 4950Asp Ala Trp Thr Asp Thr Thr Thr Leu Asp Ala Ala Tyr Trp Tyr4955 4960 4965Arg Asn Leu Arg Arg Pro Val Arg Phe Thr His Ala Val Arg Thr4970 4975 4980Leu Thr Ala Gln Gly His Arg Leu Phe Ile Glu Thr Ser Pro His4985 4990 4995Pro Thr Leu Thr Pro Ala Ile Glu Asp His Asp His Thr Thr Ala5000 5005 5010Leu Gly Thr Leu Arg Arg His Asp Asn Asp Thr His Arg Phe Leu5015 5020 5025Thr Ala Leu Ala His Ala His Thr Thr Gly His Thr Val Thr Trp
5030 5035 5040Thr Thr His Tyr Pro Thr Thr Pro His Thr Pro Ala Ile Asp Leu5045 5050 5055Pro Thr Tyr Pro Phe Gln His His His Tyr Trp Leu His Thr Pro5060 5065 5070Thr Thr Ser Thr Gly Asp Val Ser Ala Ala Gly Leu His Pro Thr5075 5080 5085Glu His Pro Leu Leu Gly Ala Thr Val Glu Leu Ala Asp Gly Asp5090 5095 5100Gly Thr Leu Leu Thr Gly Arg Leu Ser Leu His Thr His Pro Trp5105 5110 5115Leu Ala Asp His Ser Val Gly Gly Ile Val Leu Leu Pro Gly Thr5120 5125 5130Ala Leu Leu Glu Leu Ala Leu Glu Ala Gly Thr Arg Thr Gly Cys5135 5140 5145Pro His Val Gln Glu Leu Thr Leu His Thr Pro Leu Val Ile Pro5150 5155 5160Glu Thr Gly His Val Val Phe Gln Leu Thr Val Ser Ala Pro Asp5165 5170 5175Glu Thr Gly Gln Arg Pro Phe Thr Val His Phe Arg Ser Glu Ala5180 5185 5190Val Thr Gly Ala Asp Asp Pro Ala Asp Arg Thr Trp Thr Arg Cys
5195 5200 5205Ala Thr Gly Ala Leu Ser Thr Ala Ala Ala Pro Asp His Ser Glu5210 5215 5220Ala Ala Thr Trp Pro Pro Pro Ser Ala Gln Pro Leu Asp Leu Asp5225 5230 5235Gly Leu Tyr Asp Arg Met Ala Glu Ala Gly Leu Val Tyr Gly Pro5240 5245 5250Val Phe Gln Gly Leu Arg Glu Ala Trp Leu Asp Gly Glu Asp Ile5255 5260 5265Val Ala Glu Val Arg Leu Pro Gln Glu Ala Ala Ala Asp Thr Gln5270 5275 5280Gly Phe Gly Leu His Pro Ala Leu Leu Asp Ala Ala Leu His Val5285 5290 5295Thr Ala Leu Thr Ser Gln Ala Gly Thr Ala Asp Glu Asp Ala Gln5300 5305 5310Glu Arg Arg Arg Leu Pro Phe Ala Trp Ala Gly Val Ser Leu Phe5315 5320 5325Ala Arg Glu Cys Ala Ala Leu Arg Val Arg Val Ala Pro Cys Ala5330 5335 5340Pro His Pro Gly Asp Ala Val Ala Ile Thr Ala Thr Asp Glu Asp5345 5350 5355Gly Arg Pro Val Leu Ala Val Glu Ser Leu Thr Leu Arg Pro Val
5360 5365 5370Ser Pro Asp Gln Leu Arg Ala Ala Ala Pro Ala Ala Gly Arg Asp5375 5380 5385Ser Leu Phe Arg Leu Glu Trp Val Pro Val Thr Ala Ser Ala Ser5390 5395 5400Ala Ser Ala Arg Pro Thr Gly Pro Trp Ala Ala Ile Gly Thr Gly5405 5410 5415Pro Ala Val Ala Gly Leu Ala Gly His Ala Asp Leu Thr Val Tyr5420 5425 5430Ala Glu Ala Gly Asp Leu Leu Arg Asp Leu Asp Gly Gly Ala Pro5435 5440 5445Ala Pro Ala Val Val Val Leu Ser Val Thr Pro Asp Ala Asp Glu5450 5455 5460Phe Ala Thr Pro Arg Ala Ala Thr Gly Arg Ala Leu Ser Val Leu5465 5470 5475Gln Ala Trp Leu Ala Asp Glu Arg Leu Ala Asp Ser Arg Leu Val5480 5485 5490Ala Val Thr Ser Gly Ala Val Val Ala Ala Pro Gly Asp Asp Thr5495 5500 5505Val Asp Val Pro Gly Ala Ala Val Trp Gly Leu Val Arg Ser Gly5510 5515 5520Gln Ser Glu His Pro Asp Arg Ile Thr Leu Leu Asp Cys Ala Ser
5525 5530 5535Gly Ala Arg Pro Gly Pro Asp Leu Val Ala Ala Ala Leu Ala Ser5540 5545 5550Gly Glu Pro Gln Leu Ala Ala Arg Ala Gly Val Leu Tyr Thr Pro5555 5560 5565Arg Leu Ala Arg Pro His Arg Asp Ala Ser Ala Val Pro Arg Ser5570 5575 5580Leu Pro Ser His Gly Thr Val Leu Ile Thr Gly Gly Thr Gly Leu5585 5590 5595Leu Gly Gly Leu Val Ala Arg Arg Leu Val Glu Ala His Gly Val5600 5605 5610Arg Arg Leu Leu Leu Ala Gly Arg Arg Gly Pro Ala Ala Glu Gly5615 5620 5625Leu Asp Ser Leu Thr Ser Glu Leu Arg Glu Arg Gly Ala Thr Val5630 5635 5640Glu Val Ala Ala Cys Asp Ala Ala Asp Arg Thr Gln Leu Glu Ala5645 5650 5655Leu Leu Ala Gly Val Pro Glu Glu His Pro Leu Ser Ala Val Val5660 5665 5670His Ala Ala Gly Val Leu Asp Asp Gly Val Leu Thr Ser Leu Thr5675 5680 5685Asn Glu Arg Leu Gly Ala Val Leu Arg Ala Lys Ala Asp Ser Ala
5690 5695 5700Leu Leu Leu His Glu Leu Thr Gln Asp Leu Asp Leu Ser Ala Phe5705 5710 5715Val Leu Phe Ser Ser Ala Ala Gly Val Leu Gly Ser Pro Gly Gln5720 5725 5730Gly Ser Tyr Ala Ala Ala Asn Ala Val Leu Asp Ala Leu Ala His5735 5740 5745Gln Arg Ser Ala Ala Gly Leu Pro Ala Leu Ser Leu Ala Trp Gly5750 5755 5760Leu Trp Ala Glu Gly Ser Gly Met Thr Gly His Leu Asp Ala Asp5765 5770 5775Asp Arg Ser Arg Ile Asn Arg Ala Gly Met Ala Pro Leu Pro Thr5780 5785 5790Pro Asp Ala Leu Asp Leu Phe Asp Ala Ala Leu Ser Ser Asp Glu5795 5800 5805Pro Phe Leu Val Pro Ala Arg Phe Asp Leu Ser Ala Val Arg Thr5810 5815 5820Arg Thr Ala Tyr Gly Pro Leu Pro Pro Leu Leu Arg Gly Leu Val5825 5830 5835Arg Thr Ser Gly Ala His Arg Val Arg Gly Ala Val Gly Glu Ala5840 5845 5850Arg Ala Ala Gly Val Asp Glu Ala Gly Arg Leu Arg Glu Arg Leu
5855 5860 5865Ala Arg Gln Ser Asp Ala Glu Arg Arg Asn Thr Leu Leu Arg Leu5870 5875 5880Val Gln Ser Asn Val Ala Ala Val Leu Gly His Arg Gly Thr Gly5885 5890 5895Thr Val Ala Glu Thr Arg Ala Phe Arg Glu Leu Gly Phe Asp Ser5900 5905 5910Leu Thr Ala Val Glu Leu Arg Asn Arg Leu Lys Val Ala Thr Gly5915 5920 5925Leu Ala Leu Arg Ala Thr Val Ala Phe Asp Phe Pro Thr Pro Ala5930 5935 5940Ala Leu Ala Glu His Leu Gly Ala Arg Leu Leu Pro Pro Asp Gly5945 5950 5955Ala Val Ser Glu Ala Val Gly Glu Lys Glu Leu Arg Gly Leu Leu5960 5965 5970Thr Ser Ile Pro Ile Gly Arg Leu Arg Glu Ala Gly Leu Ile Asp5975 5980 5985Arg Leu Leu Ala Leu Ala Ala Ala Ala Pro Asp Ser Ala Asp Gln5990 5995 6000Thr Ala Glu Gln Pro Ser Arg Ser Val Ser Val Glu Asp Ile Asp6005 6010 6015Ala Met Asp Val Asp Ser Leu Ile Gly Leu Ala His Asp Thr Gly
6020 6025 6030Thr Asp Ser Gly His Ala Pro Cys Glu Gly6035 6040<210>8<211>284<212>PRT<213>细菌<400>8Met Thr Lys Ala Pro His Gln Gly Ser Pro Thr Pro Ala Asp Val Gly1 5 10 15Asp Tyr Tyr Asp Arg Met Thr Ser Leu Leu Asn Arg Ala Leu Gly Gly20 25 30Asn Thr His Leu Gly Tyr Trp Pro His Pro Asp Asp Gly Ser Thr Leu35 40 45Gly Gln Ala Ser Asp Arg Leu Thr Asp His Met Ile Gly Lys Leu Arg50 55 60Glu His Thr Gly Arg Pro Val Arg Arg Val Leu Asp Val Gly Cys Gly65 70 75 80Ser Gly Arg Pro Ala Leu Arg Leu Ala His Ser Glu Pro Val Asp Ile85 90 95Val Gly Ile Thr Ile Ser Pro Arg Gln Val Glu Leu Ala Thr Ala Leu100 105 110Ala Glu Arg Ser Gly Leu Ala Asn Arg Val Arg Phe Glu Cys Ala Asp115 120 125
Ala Met Asp Leu Pro Phe Pro Asp Ala Ser Phe Asp Ala Val Trp Ala130 135 140Leu Glu Cys Leu Leu His Met Pro Asp Pro Ala Arg Val Phe Gln Glu145 150 155 160Met Ala Arg Val Leu Arg Pro Gly Gly Arg Leu Ala Ala Met Asp Val165 170 175Thr Leu Arg Ala Ser Gln Pro Thr Gly Ala Asp Trp Ser Ser Ser Glu180 185 190Leu Ala Val Pro Ser Leu Ile Pro Ile Thr Ala Tyr Ala Gly Met Ile195 200 205Ser Asp Ala Gly Leu Arg Leu Thr Glu Leu Thr Asp Ile Gly Glu His210 215 220Val Ile Ala Pro Ser Tyr Ser Ala Met Gly Asp Asp Val Arg Ala Asn225 230 235 240Ala His Ala Tyr Ala Glu Ala Leu Glu Met Thr Ala Asp Asp Leu Glu245 250 255Thr Phe Val Gly Lys Cys Ser Gln Trp Tyr Thr Glu Asp Ile Gly Tyr260 265 270Val Val Leu Thr Ala Pro Cys Gln Arg Ala Glu Val275 280<210>9<211>468
<212>PRT<213>细菌<400>9Val Ser Ser Pro Pro Ser Thr Ile Pro Glu Ala Pro Gly Ala Trp Pro1 5 10 15Val Leu Gly His Leu Pro Ala Leu Leu Arg Asp Pro Leu Gly Phe Leu20 25 30Ser Ala Val Thr Glu Arg Gly Asp Leu Phe Arg Ile Arg Leu Gly His35 40 45Asn Thr Val Tyr Leu Ala Thr His Pro Glu Ile Val Arg Thr Met Leu50 55 60Val Ser Gly Ala Ala Asp Phe Thr Arg Ser Lys Gly Ala Ala Gly Ala65 70 75 80Ser Arg Phe Ile Gly Pro Ile Leu Val Ala Val Ser Gly Asp Ser His85 90 95Arg Arg Gln Arg Arg Met Met Gln Pro Gly Phe His Arg Gly Lys Leu100 105 110Asp His Tyr Val Ile Ser Met Ser Ala Ala Ala Glu Glu Thr Ala Asp115 120 125Ser Trp Arg Pro Gly Gln Val Val Asp Val Pro Lys Met Ala Ser Asp130 135 140Leu Ser Leu Ala Met Ile Thr Lys Ala Leu Phe Gln Ser Asp Leu Gly145 150 155 160
Ala Ala Ala Glu Ala Glu Leu Arg Thr Thr Gly His Asp Ile Leu Lys165 170 175Val Ala Arg Leu Ser Ala Leu Ala Pro Gln Leu Tyr Thr Ser Leu Pro180 185 190Thr Ala Ala Lys Arg His Met Gly Arg Thr Ser Ala Ala Ile Arg Glu195 200 205Ala Val Thr Ala Tyr Arg Ala Asp Gly Arg Asp His Gly Asp Leu Leu210 215 220Ser Thr Met Leu Arg Ala Arg Asp Ala Glu Gly Asn Thr Met Thr Asp225 230 235 240Asp Glu Val His Asn Glu Ile Met Gly Leu Ala Val Ala Gly Ile Gly245 250 255Gly Pro Ala Ala Leu Thr Ala Trp Ile Phe His Glu Leu Ala His Asp260 265 270His Leu Ile Glu Gln Arg Leu His Ala Glu Ile Asp Thr Val Leu Gly275 280 285Gly Arg Leu Pro Thr Ser Ala Asp Leu Pro Arg Leu Pro Tyr Thr Gln290 295 300Arg Leu Val Lys Glu Ala Leu Arg Lys Tyr Pro Gly Trp Val Gly Ser305 310 315 320Arg Arg Thr Val Arg Pro Val Arg Leu Gly Glu His Glu Leu Pro Ala325 330 335
Asp Val Glu Ile Met Tyr Ser Ser Tyr Ala Leu Gln Arg Asp Pro Arg340 345 350Trp Tyr Arg Asp Pro Glu Lys Leu Asp Pro Asp Arg Trp Glu Ser Lys355 360 365Glu Thr Thr Arg Asp Val Pro Lys Gly Ala Trp Val Pro Phe Ala Leu370 375 380Gly Thr Tyr Lys Cys Ile Gly Asp Asn Phe Ala Leu Met Glu Thr Ala385 390 395 400Val Ala Val Ala Val Ile Ala Ser Arg Trp Arg Leu Arg Pro Leu Lys405 410 415Gly Asp Arg Val Arg Pro Val Ala Lys Ala Thr His Val Phe Pro Asp420 425 430Arg Leu Arg Met Ile Ala Glu Pro Arg Thr Pro Ala Ile Pro Arg Gly435 440 445His Ala Pro Ala Asp Ala Ser Leu Glu Ala Ala Ala Arg Pro Lys Glu450 455 460Leu Pro Glu Pro465<210>10<211>5674<212>PRT<213>细菌<400>10
Met Ala Thr Pro Ser Glu Lys Leu Val Glu Ala Leu Arg Ala Ser Leu1 5 10 15Lys Ala Asn Glu Ala Leu Arg Arg Arg Asn Gln Gln Leu Thr Ala Ala20 25 30Val Glu Ala Ala Gln Glu Pro Leu Ala Ile Val Gly Met Ala Cys Arg35 40 45Phe Pro Gly Gly Val Arg Ser Pro Glu Glu Leu Trp Gly Leu Val Ala50 55 60Ser Gly Gly Asp Ala Ile Gly Glu Phe Pro Ala Asp Arg Gly Trp Asp65 70 75 80Leu Ala Gly Leu Phe Asp Pro Asp Pro Glu Arg Ala Gly Ala Ser Tyr85 90 95Thr Arg His Gly Gly Phe Leu Tyr Asp Ala Gly Gln Phe Asp Ala Glu100 105 110Leu Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln115 120 125Arg Leu Leu Leu Glu Thr Ser Trp Glu Val Phe Glu Arg Ala Gly Ile130 135 140Asp Pro Ser Ser Val Arg Gly Ala Arg Ala Gly Val Phe Thr Gly Met145 150 155 160Met Tyr His Asp Tyr Ala Ser Arg Leu Ala Thr Ile Pro Glu Gly Phe165 170 175
Glu Gly Tyr Ile Gly Asn Gly Ser Gly Gly Ala Val Ala Ser Gly Arg180 185 190Val Ala Tyr Thr Leu Gly Leu Glu Gly Pro Ala Val Thr Val Asp Thr195 200 205Ala Cys Ser Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ser Leu210 215 220Arg Thr Gly Glu Cys Asp Leu Ala Leu Ala Gly Gly Val Thr Val Met225 230 235 240Ser Thr Pro Leu Leu Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ser245 250 255Val Asp Gly Arg Cys Lys Ser Phe Ala Ala Ala Ala Asp Gly Thr Gly260 265 270Met Gly Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu Ser Asp Ala275 280 285Glu Arg Asn Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val290 295 300Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser305 310 315 320Gln Glu Arg Val Ile Arg Glu Ala Leu Ala Asn Ala Gly Leu Thr Val325 330 335Ala Asp Val Asp Ala Val Glu Gly His Gly Thr Gly Thr Arg Leu Gly340 345 350
Asp Pro Ile Glu Ala Gln Ala Leu Leu Asp Thr Tyr Gly Gln Glu Arg355 360 365Ser Gly Glu Gln Pro Leu Trp Leu Gly Ser Val Lys Ser Asn Ile Gly370 375 380His Ala Gln Ala Ala Ala Gly Val Gly Gly Ile Ile Lys Met Val Met385 390 395 400Ala Leu Arg His Glu Ser Leu Pro Arg Thr Leu His Val Asp Glu Pro405 410 415Ser Pro Gln Val Asp Trp Ser Ser Gly Ala Val Ser Leu Leu Ser Glu420 425 430Ala Arg Pro Trp Pro Arg Arg Glu Asp Arg Pro Arg Arg Ala Gly Val435 440 445Ser Ser Phe Gly Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Glu450 455 460Ala Pro Ala Arg Arg Pro Gly Glu Ala Ala Val Glu Asp Gly Ala Pro465 470 475 480Val Pro Trp Val Val Ser Ala Arg Ser Gly Ala Ala Leu Arg Ala Gln485 490 495Ala Met Val Leu Arg Glu Phe Leu Arg Gly Pro Gly Thr Asp Ala Gly500 505 510Val Arg Asp Ile Gly Ala Glu Leu Ala Arg Gly Arg Ala Val Leu Glu515 520 525
His Arg Ala Val Ile Val Ala Arg Glu Arg Ala Glu Phe Glu Gly Ala530 535 540Leu Glu Ala Leu Ala Ser Gly Glu Pro His Pro Ala Leu Ile Glu Asp545 550 555 560Ala Thr Gly Ser His Ser His Ser Gly Gly Gly Val Val Phe Val Phe565 570 575Pro Gly Gln Gly Gly Gln Trp Ala Gly Met Gly Leu Asp Leu Leu Thr580 585 590Thr Ser Gly Val Phe Ala Asp His Ile Gly Ala Cys Glu Arg Ala Leu595 600 605Ala Pro Trp Val Glu Trp Ser Leu Thr Glu Met Leu His Arg Glu Ala610 615 620Glu Asp Pro Val Trp Glu Arg Ala Asp Val Val Gln Pro Val Leu Phe625 630 635 640Ser Val Met Val Ser Leu Ala Ala Leu Trp Arg Ser Tyr Gly Ile Glu645 650 655Pro Asp Ala Val Val Gly His Ser Gln Gly Glu Ile Ala Ala Ala His660 665 670Val Cys Gly Ala Leu Thr Leu Glu Asp Ala Ala Lys Val Val Ala Leu675 680 685Arg Ser Arg Ala Leu Ala Ala Leu Arg Gly His Gly Gly Met Ala Ser690 695 700
Leu Ala Leu Thr Gly Thr Glu Ala Glu Asp Leu Ile Thr Thr His Trp705 710 715 720Pro Gly Arg Leu Trp Thr Ala Ala Phe Asn Gly Pro Arg Ala Thr Thr725 730 735Val Ser Gly Asp Thr Asp Ala Leu Asp Glu Leu Leu Thr His Cys Thr740 745 750Glu Thr Gly Val Arg Ala Arg Arg Ile Pro Val Asp Tyr Ala Ser His755 760 765Cys Pro His Thr Glu Thr Ile Glu His Asp Leu Leu His Met Leu His770 775 780Gly Ile Thr Pro Gln Pro Gly Ser Ile Pro Phe Tyr Ser Thr Val Glu785 790 795 800Asp Ala Trp Thr Asp Thr Thr Thr Leu Asp Ala Ala Tyr Trp Tyr Arg805 810 815Asn Leu Arg Arg Pro Val Arg Phe Thr His Ala Val Arg Thr Leu Thr820 825 830Ala Gln Gly His Arg Leu Phe Ile Glu Thr Ser Pro His Pro Thr Leu835 840 845Thr Pro Ala Ile Glu Asp His Asp His Thr Thr Ala Leu Gly Thr Leu850 855 860Arg Arg His Asp Asn Asp Thr His Arg Phe Leu Thr Ala Leu Ala His865 870 875 880
Ala His Thr Thr Gly His Thr Val Thr Trp Thr Thr His Tyr Pro Thr885 890 895Thr Pro His Thr Pro Ala Ile Asp Leu Pro Thr Tyr Pro Phe Gln His900 905 910His His Tyr Trp Leu His Thr Pro Thr Thr Ser Thr Gly Asp Val Ser915 920 925Ala Ala Gly Leu Gln Arg Pro Asp His Pro Leu Leu Gly Ala Val Met930 935 940Glu Leu Ala Asp Gly Asp Gly Ile Val Leu Thr Gly Arg Leu Ser Leu945 950 955 960His Thr His Pro Trp Leu Ala Asp His Ser Val Gly Gly Val Val Leu965 970 975Leu Pro Gly Thr Ala Leu Leu Glu Leu Ala Phe Gln Ala Gly Leu Arg980 985 990Ala Gly Cys Pro Gly Val Asp Glu Leu Thr Leu His Ala Pro Leu Val995 1000 1005Val Pro Glu Ser Gly His Val Val Val Gln Val Ser Val Ser Val1010 1015 1020Pro Asp Glu Ala Gly Arg Arg Gly Val Ser Val Tyr Gly Arg Leu1025 1030 1035Val Glu Asp Gly Gly Leu Glu Gly Glu Trp Thr Arg His Ala Glu1040 1045 1050
Gly Val Val Cys Pro Ser Val Pro Gly Glu Ser Val Val Val Glu1055 1060 1065Pro Val Ala Asp Gly Val Trp Pro Pro Ser Gly Ala Gln Pro Val1070 1075 1080Asp Leu Asp Glu Phe Tyr Gly Arg Leu Ala Gly Gly Gly Phe Val1085 1090 1095Tyr Gly Pro Val Phe Gln Gly Leu Cys Ala Ala Trp Arg Asp Gly1100 1105 1110Asp Asp Val Val Ala Glu Val Arg Leu Pro Asp Glu Gly Leu Ala1115 1120 1125Asp Val Ala Gly Phe Gly Val His Pro Ala Leu Leu Asp Ala Ala1130 1135 1140Val Gln Thr Val Thr Leu Leu Leu Pro Glu Asp Gln Glu Ala Gly1145 1150 1155Leu Leu Pro Tyr Thr Trp Asn Gly Ala Ser Leu His Ala Arg Gly1160 1165 1170Ala Arg Ala Leu Arg Val Arg Val Thr Ser Val Asp Ala Ala Gly1175 1180 1185Thr Thr Val Ser Leu Arg Val Ala Asp Glu Thr Gly Ala Leu Val1190 1195 1200Leu Ala Leu Glu Ser Leu Val Leu Arg Pro Val Pro Leu Glu Gly1205 1210 1215
Leu Gly Ala Gly Val Arg Arg Gly Ser Leu Phe Glu Leu Gly Trp1220 1225 1230Val Pro Val Glu Gly Val Pro Ala Ser Leu Ala Gly Gly Gly Gly1235 1240 1245Glu Leu Val Val Trp Glu Cys Pro Gly Gly Gly Val Ala Glu Val1250 1255 1260Thr Ala Ala Ala Leu Gly Val Val Arg Glu Trp Leu Ala Asp Glu1265 1270 1275Arg Glu Gly Asp Ala Arg Leu Val Val Val Thr Arg Gly Ala Val1280 1285 1290Ala Val Asp Ala Gly Glu Pro Val Arg Asp Val Ala Gly Ala Ala1295 1300 1305Val Trp Gly Leu Val Arg Ser Ala Gln Ser Glu His Pro Asp Arg1310 1315 1320Phe Val Leu Leu Asp Leu Asp Pro Gly Thr Gly Val Glu Thr Val1325 1330 1335Val Asp Ala Asp Glu Asp Met Gly Ala Gly Val Gly Ala Gly Val1340 1345 1350Asp Val Ala Gly Phe Val Ala Cys Gly Glu Ala Gln Val Ala Val1355 1360 1365Arg Gly Gly Val Val Arg Val Pro Arg Leu Glu Arg Leu Glu Arg1370 1375 1380
Trp Gly Arg Leu Gly Gly Ala Gly Glu Gly Leu Ser Leu Pro Gly1385 1390 1395Gly Val Gly Trp Arg Leu Asp Gly Gly Gly Ser Gly Leu Leu Glu1400 1405 1410Gly Val Gly Val Val Ala Ser Asp Ala Ala Gly Val Val Leu Gly1415 1420 1425Arg Gly Gln Val Arg Val Ala Val Arg Ala Ala Gly Val Asn Phe1430 1435 1440Arg Asp Val Leu Val Ala Leu Gly Met Val Pro Gly Gln Val Gly1445 1450 1455Val Gly Ser Glu Gly Ala Gly Val Val Val Glu Val Gly Pro Gly1460 1465 1470Val Glu Gly Leu Val Val Gly Asp Arg Val Phe Gly Val Phe Gly1475 1480 1485Asp Ala Phe Ala Pro Val Val Val Ala Gln Glu Val Leu Leu Ala1490 1495 1500Arg Ile Pro Glu Gly Trp Ser Phe Ala Gln Ala Ala Ser Val Pro1505 1510 1515Val Val Phe Ala Thr Ala Tyr Leu Gly Leu Val Asp Leu Ala Gly1520 1525 1530Val Arg Arg Gly Glu Ser Val Leu Val His Ala Ala Ala Gly Gly1535 1540 1545
Val Gly Thr Ala Ala Val Gln Leu Ala Arg His Leu Gly Ala Glu1550 1555 1560Val Tyr Ala Thr Ala Ser Glu Ala Lys Trp Ala Arg Leu Arg Ala1565 1570 1575Ala Gly Val Ala Pro Gln Arg Ile Ala Ser Ser Arg Ser Val Glu1580 1585 1590Phe Glu Ser Arg Phe Arg Arg Ala Ser Gly Gly Arg Gly Val Asp1595 1600 1605Val Val Leu Asn Cys Leu Ala Gly Glu Tyr Thr Asp Ala Ser Leu1610 1615 1620Arg Leu Cys Ser Pro Gln Gly Gly Arg Phe Leu Glu Leu Gly Lys1625 1630 1635Thr Asp Ile Arg Asp Ala Gly Glu Val Ala Ala Arg Phe Pro Gly1640 1645 1650Val Ser Tyr Arg Ala Tyr Asp Leu Met Asp Ala Gly Ala Gln Arg1655 1660 1665Val Gly Glu Ile Leu His Thr Val Val Asp Leu Phe Arg Arg Gly1670 1675 1680Val Leu Glu Pro Leu Pro Val Thr Ala Trp Asp Val Arg Gln Ala1685 1690 1695Arg Gln Ala Leu Arg Ser Met Arg Ser Gly Leu His Val Gly Lys1700 1705 1710
Asn Val Leu Thr Leu Pro Val Pro Leu Asp Ala Glu Gly Thr Val1715 1720 1725Leu Val Thr Gly Gly Thr Gly Thr Leu Gly Ala Ala Val Ala Arg1730 1735 1740His Leu Ala Ala Gly His Gly Val Arg His Leu Leu Leu Val Ser1745 1750 1755Arg Arg Gly Met Ala Ala Ala Gly Ala Glu Glu Leu Cys Ala Glu1760 1765 1770Leu Gly Gln Ala Gly Val Ser Val Ser Val Ala Ala Cys Asp Val1775 1780 1785Ala Asp Arg Ala Gln Val Ala Ala Leu Leu Glu Gln Val Pro Ala1790 1795 1800Glu His Pro Leu Thr Ala Val Val His Thr Ala Gly Val Leu Asp1805 1810 1815Asp Ala Thr Val Thr Cys Leu Asp Arg Glu Lys Ile Asp Ala Val1820 1825 1830Val Gly Ala Lys Val Asp Gly Ala Leu His Leu His Glu Leu Thr1835 1840 1845Ala Gly Met Asp Leu Ser Ala Phe Val Leu Phe Ser Ser Ala Ala1850 1855 1860Gly Val Leu Gly Ser Pro Gly Gln Gly Asn Tyr Ala Ala Ala Asn1865 1870 1875
Ala Ala Leu Asp Ala Leu Ala His Gln Arg Arg Ala Ala Gly Leu1880 1885 1890Pro Ala Leu Ser Leu Ala Trp Gly Leu Trp Glu Glu Ala Ser Gly1895 1900 1905Met Thr Gly His Leu Asp Ala Gly Asp Arg His Arg Ile Thr Arg1910 1915 1920Ser Gly Leu His Pro Leu Thr Thr Pro Asp Ala Leu Ala Leu Leu1925 1930 1935Asp Thr Ala Leu Ala Thr Gly Arg Pro Ala Leu Leu Pro Ala Asp1940 1945 1950Leu Arg Pro Thr His Pro Ala Pro Pro Leu Leu Glu His Leu Ala1955 1960 1965Pro Ala Arg Thr Ser Pro Arg Thr Ala His Thr Gly Thr Ser Ala1970 1975 1980Gly Ala Gly Gln Asp Val Ser Leu Ala Asp Arg Leu Ala Thr Leu1985 1990 1995Thr Ser Glu Gln Arg His Ala Thr Leu Leu Ala Leu Ala Arg Thr2000 2005 2010His Ile Ala Ala Val Leu Gly His Pro Thr Pro Asp Thr Ile Asp2015 2020 2025Pro Glu Arg Thr Phe Arg Asp Leu Gly Phe Asp Ser Leu Thr Ala2030 2035 2040
Val Glu Leu Arg Asn Arg Leu Thr Arg Ala Thr Gly Leu Arg Leu2045 2050 2055Pro Thr Thr Leu Ala Phe Asp His Pro Thr Pro Thr Ala Leu Thr2060 2065 2070His His Leu Thr Thr Leu Leu Asn Pro Asn Asp Thr Lys Thr Pro2075 2080 2085Ser Ala Pro Ala Ala Ala Glu Pro Lys Ala Gly Gln His Glu Pro2090 2095 2100Ile Ala Ile Ile Gly Val Gly Cys Arg Tyr Pro Gly Gly Val Ala2105 2110 2115Ser Ala Glu Asp Leu Trp Gln Leu Val Ala Ser Gly Gly Asp Ala2120 2125 2130Val Gly Glu Phe Pro Ala Asp Arg Gly Trp Asp Val Glu Ala Leu2135 2140 2145Tyr Asp Pro Glu Pro Gly Gln Arg Gly Thr Ser Tyr Thr Arg His2150 2155 2160Gly Gly Phe Leu Tyr Asp Ala Gly Glu Phe Asp Ala Gly Phe Phe2165 2170 2175Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln Arg2180 2185 2190Leu Leu Leu Glu Thr Thr Trp Glu Ala Phe Glu Arg Ala Gly Ile2195 2200 2205
Asp Pro Gly Ala Val Arg Gly Ser Gln Thr Gly Val Phe Ala Gly2210 2215 2220Val Met Pro Gln Glu Tyr Ala Ser Arg Ser Arg His His Val Ala2225 2230 2235Ala Asp Val Asp Gly Tyr Val Leu Thr Gly Thr Ser Gly Ser Val2240 2245 2250Ala Ser Gly Arg Val Ala Tyr Thr Phe Gly Leu Glu Gly Pro Ala2255 2260 2265Val Ser Val Asp Thr Ala Cys Ser Ser Ser Leu Val Ala Leu His2270 2275 2280Leu Ala Cys Gln Ala Leu Arg Ser Gly Glu Cys Thr Met Ala Leu2285 2290 2295Ala Gly Gly Ala Thr Val Met Ser Thr Pro Thr Ala Phe Leu Glu2300 2305 2310Phe Ser Arg Gln Arg Gly Leu Ala Ala Asp Gly Arg Cys Lys Ala2315 2320 2325Phe Ser Ala Ser Ala Asp Gly Thr Gly Trp Ser Glu Gly Ala Gly2330 2335 2340Met Leu Leu Leu Glu Arg Leu Ser Asp Ala Glu Arg Asn Gly His2345 2350 2355Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly2360 2365 2370
Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Gln Arg2375 2380 2385Val Ile Arg Gln Ala Leu Ala Asn Ala Asn Leu Ser Ala Val Asp2390 2395 2400Val Asp Ala Val Glu Ala His Gly Thr Gly Thr Lys Leu Gly Asp2405 2410 2415Pro Ile Glu Ala Gln Ala Leu Leu Ala Thr Tyr Gly Gln Glu His2420 2425 2430His Pro Asp Gln Pro Leu Trp Leu Gly Ser Leu Lys Ser Asn Ile2435 2440 2445Gly His Ala Gln Ala Ala Ala Gly Val Gly Gly Ile Ile Lys Met2450 2455 2460Val Met Ala Leu Arg His Glu Ser Leu Pro Arg Thr Leu His Val2465 2470 2475Asp Glu Pro Ser Pro Gln Val Asp Trp Ser Ser Gly Ala Val Ser2480 2485 2490Leu Leu Thr Glu Ala Arg Pro Trp Pro Arg Arg Glu Asp Arg Pro2495 2500 2505Arg Arg Ala Gly Ile Ser Ser Phe Gly Val Ser Gly Thr Asn Ala2510 2515 2520His Val Ile Leu Glu Glu Ala Pro Ala Arg Ala Glu Val Glu Ala2525 2530 2535
Val Glu Ala Ala Pro Ala Gly Val Glu Thr Ala Ala Ala Ala Ala2540 2545 2550Val Val Val Glu Thr Asp Gly Ala Gly Arg Val Ser Ala Asp Val2555 2560 2565Pro Leu Val Trp Val Val Ser Gly Lys Ser Gln Ala Ala Leu Arg2570 2575 2580Ala Gln Ala Ala Ala Leu His Ala His Val Leu Asp His Pro Glu2585 2590 2595Gln Asp Ala Ala Asp Ile Gly Tyr Ser Leu Ala Thr Thr Arg Ala2600 2605 2610Leu Phe Asp His Arg Ala Thr Leu Ile Ala Pro Asp Arg Asp Thr2615 2620 2625Leu Leu Asp Ala Leu Thr Ala Leu Ala Asp Gly Arg Thr His Pro2630 2635 2640His Leu Ile Pro Thr Pro Pro Thr Glu Pro Gly His Thr His Lys2645 2650 2655Ile Ala Phe Leu Cys Ser Gly Gln Gly Thr Gln Arg Pro Gly Met2660 2665 2670Ala Thr Gly Leu Tyr His Thr Tyr Pro Ala Phe Ala Asp Ala Leu2675 2680 2685Asp Glu Thr Cys Ala His Phe Asp Pro His Leu Asp His Pro Leu2690 2695 2700
Arg Asp Leu Leu Leu Asn His Asp Pro Thr Asp Leu Leu Thr His2705 2710 2715Thr Leu Tyr Ala Gln Pro Ala Leu Phe Thr Leu Gln Lys Ala Leu2720 2725 2730His His Leu Ile Thr Glu Thr Tyr Gly Ile Thr Pro His Tyr Leu2735 2740 2745Ala Gly His Ser Leu Gly Glu Ile Thr Ala Ala His Leu Ala Gly2750 2755 2760Ile Leu Thr Leu Pro Asp Ala Thr His Leu Ile Thr Thr Arg Ala2765 2770 2775Arg Leu Met Gln Thr Met Pro Pro Gly Thr Met Thr Thr Leu His2780 2785 2790Thr Thr Pro Glu His Ile Gln Pro Leu Leu Asp Gln His Pro Gly2795 2800 2805Lys Ala Thr Ile Ala Ala Val Asn Ser Pro His Ser Leu Val Ile2810 2815 2820Ser Gly Asp Pro Asp Thr Ile His His Ile Thr Thr Thr Cys His2825 2830 2835Thr Gln Gly Ile Thr Thr Lys Pro Leu Thr Thr Asn His Ala Phe2840 2845 2850His Ser Pro His Thr Asp Thr Ile Leu Glu Gln Leu Asp Thr Thr2855 2860 2865
Thr His Thr Leu Thr Tyr His Pro Pro His Thr Pro Leu Ile Thr2870 2875 2880Ser Thr Pro Gly Asp Pro Leu Thr Pro His Tyr Trp Thr His Gln2885 2890 2895Thr Arg Gln Pro Val His Trp Thr Asp Thr Ile His Thr Leu His2900 2905 2910Thr Asn Gly Val Thr Thr Tyr Ile Glu Leu Gly Pro Asp His Thr2915 2920 2925Leu Thr Thr Leu Thr His His Asn Leu Pro His His Gln Pro Thr2930 2935 2940Ala Ile Thr Leu Thr His Pro His His Asn Pro Thr His His Leu2945 2950 2955Leu Thr Ala Leu Ala His Thr Pro Thr Thr Trp His Thr His His2960 2965 2970His Thr His Thr Asn Pro His Pro His Thr Ile Pro Asp Leu Pro2975 2980 2985Thr Tyr Pro Phe Gln Arg Arg His Tyr Trp Leu Gln Ala Thr Pro2990 2995 3000Gly Ala Gly Ala Gly Asp Val Ser Ala Ala Gly Leu Gln Arg Pro3005 3010 3015Asp His Pro Leu Leu Gly Ala Val Met Glu Leu Ala Asp Gly Asp3020 3025 3030
Gly Ile Val Leu Thr Gly Ser Leu Ser Leu Arg Thr His Thr Trp3035 3040 3045Leu Ala Asp His Ser Val Gly Gly Ile Val Leu Leu Pro Gly Thr3050 3055 3060Ala Leu Leu Asp Leu Ala Phe Gln Ala Gly Leu Arg Thr Gly Cys3065 3070 3075Pro Arg Val Asp Glu Leu Thr Leu His Ala Pro Leu Val Ile Pro3080 3085 3090Glu Ser Gly His Val Val Val Gln Val Ser Val Ser Val Pro Asp3095 3100 3105Glu Ala Gly Arg Arg Ala Val Asn Val Tyr Ala Arg Pro Ala Gly3110 3115 3120Asp Glu Glu Thr Asp Gly Glu Trp Thr Arg His Ala Glu Gly Val3125 3130 3135Leu Ser Pro Ser Thr Glu Asp Asp Pro Asn Ala Glu Ala Ala Ala3140 3145 3150Ala Gly Glu Trp Pro Pro Pro Gly Ala Arg Pro Val Val Leu Asp3155 3160 3165Gly Leu Tyr Asp Arg Leu Ala Gly Gly Gly Phe Val Tyr Gly Pro3170 3175 3180Val Phe Gln Gly Leu Cys Ala Ala Trp Arg Asp Gly Asp Asp Val3185 3190 3195
Val Ala Glu Val Arg Leu Pro Asp Glu Gly Leu Ala Asp Val Ala3200 3205 3210Gly Phe Gly Val His Pro Ala Leu Leu Asp Ala Ala Val Gln Ser3215 3220 3225Val Thr Leu Leu Leu Ala Asp Gln Gln Gln Ala Gly Leu Val Pro3230 3235 3240His Thr Trp Asn Gly Val Ser Leu His Ala Arg Gly Ala Thr Val3245 3250 3255Leu Arg Leu Arg Met Thr Pro Thr Asp Ala Thr Ser Thr Ala Val3260 3265 3270Arg Leu His Ala Thr Asp Glu Thr Gly Ala Pro Val Leu Thr Leu3275 3280 3285Glu Ser Leu Leu Met Arg Pro Val Pro Leu Glu Gly Leu Gly Ala3290 3295 3300Arg Val Arg Arg Gly Ser Leu Phe Glu Leu Gly Trp Val Pro Val3305 3310 3315Glu Gly Val Pro Ala Ser Val Ala Gly Gly Gly Gly Glu Leu Val3320 3325 3330Ala Trp Glu Cys Pro Gly Gly Gly Val Ala Glu Val Thr Ala Ala3335 3340 3345Ala Leu Gly Val Val Arg Glu Trp Leu Ala Asp Glu Arg Glu Gly3350 3355 3360
Asp Ala Arg Leu Val Val Val Thr Arg Gly Ala Val Ala Val Asp3365 3370 3375Ala Gly Glu Pro Val Arg Asp Val Ala Gly Ala Ala Val Trp Gly3380 3385 3390Leu Val Arg Ser Ala Gln Ser Glu His Pro Asp Arg Phe Val Leu3395 3400 3405Leu Asp Leu Asp Pro Asp Thr Lys Thr Asp Pro Asp Thr Asp Thr3410 3415 3420Asp Thr Asp Thr Asp Gly Asp Thr Asp Val Ser Ala Asp Ala Lys3425 3430 3435Val Gly Thr Gly Ala Gly Leu Asp Asp Ala Ala Val Ala Ser Ala3440 3445 3450Leu Ala Arg Gly Glu Ser Gln Leu Ala Val Arg Asp Gly Val Val3455 3460 3465Arg Val Pro Arg Leu Lys Arg Val Pro Pro Leu Ser Glu Ser Ser3470 3475 3480Asp Ala Val Arg Phe Asp Ala Glu Gly Thr Val Leu Val Thr Gly3485 3490 3495Gly Thr Gly Thr Leu Gly Ala Val Val Ala Arg His Leu Ala Ala3500 3505 3510Gly His Gly Val Arg His Leu Leu Leu Val Ser Arg Arg Gly Met3515 3520 3525
Ala Ala Thr Gly Ala Glu Glu Leu Cys Ala Glu Leu Gly Gly Ala3530 3535 3540Gly Val Ser Val Ser Val Ala Ala Cys Asp Val Ala Asp Arg Ala3545 3550 3555Gln Val Ala Ala Leu Leu Glu Gln Val Pro Ala Glu His Pro Leu3560 3565 3570Thr Ala Val Val His Thr Ala Gly Val Leu Asp Asp Ala Thr Val3575 3580 3585Thr Cys Leu Asp Arg Glu Lys Ile Asp Ala Val Val Gly Ala Lys3590 3595 3600Val Asp Gly Ala Leu His Leu His Glu Leu Thr Ala Gly Met Asp3605 3610 3615Leu Ser Ala Phe Val Leu Phe Ser Ser Ala Ala Gly Val Leu Gly3620 3625 3630Ser Pro Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Ala Leu Asp3635 3640 3645Ala Leu Ala His Gln Arg Arg Ala Ala Gly Leu Pro Ala Leu Ser3650 3655 3660Leu Ala Trp Gly Leu Trp Glu Glu Thr Ser Gly Met Thr Gly His3665 3670 3675Leu Asp Ala Gly Asp Arg His Arg Ile Thr Arg Ser Gly Leu His3680 3685 3690
Pro Leu Thr Thr Pro Asp Ala Leu Ala Leu Leu Asp Thr Ala Leu3695 3700 3705Ala Ala Gly Arg Pro Ala Leu Leu Pro Ala Asp Leu Arg Pro Thr3710 3715 3720His Pro Ala Pro Pro Leu Leu Glu His Leu Ala Pro Ala Arg Thr3725 3730 3735Ser His Arg Thr Thr Leu Pro Thr Thr Asp Ser Gly Ala Ser Leu3740 3745 3750Arg Ala Arg Leu Ala Gly Arg Thr Pro Glu Gln Gln Tyr Gln Ala3755 3760 3765Leu Leu Gly Leu Val Arg Ser His Val Ala Thr Val Leu Gly His3770 3775 3780Gln Ala Pro Glu Ala Ile Pro Val Asp Ser Ala Phe Arg Asp Leu3785 3790 3795Gly Phe Asp Ser Leu Thr Ala Val Asp Leu Arg Asn Arg Leu Ser3800 3805 3810Ala Glu Thr Gly Leu Arg Leu Pro Ala Ser Leu Val Phe Asp Gln3815 3820 3825Pro Ser Pro Ala Ala Val Ala Arg Leu Leu Arg Thr Glu Leu Leu3830 3835 3840Gly Asp Asp Ala Ala Asp Ser Thr Ser Pro Tyr Ala Glu Thr Thr3845 3850 3855
Ala Val Gly Ser Asp Glu Pro Leu Ala Ile Val Gly Met Ala Cys3860 3865 3870Arg Phe Pro Gly Gly Val Arg Ser Pro Glu Glu Leu Trp Gly Leu3875 3880 3885Val Ala Ser Gly Gly Asp Ala Ile Gly Glu Phe Pro Ala Asp Arg3890 3895 3900Gly Trp Asp Leu Ala Gly Leu Phe Asp Pro Asp Pro Glu Arg Ala3905 3910 3915Gly Ala Ser Tyr Thr Arg His Gly Gly Phe Leu Tyr Asp Ala Gly3920 3925 3930Gln Phe Asp Ala Glu Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu3935 3940 3945Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Thr Val Trp Glu3950 3955 3960Thr Leu Glu His Ala Gly Ile Asp Pro Ala Ala Val Arg Gly Ser3965 3970 3975Arg Thr Gly Val Phe Ala Gly Val Met Tyr His Asp Tyr Ala Ala3980 3985 3990Arg Leu Thr Ala Val Pro Glu Gly Ala Glu Gly Tyr Ile Gly Asn3995 4000 4005Gly Asn Ala Gly Ser Val Val Ser Gly Arg Val Ala Tyr Thr Phe4010 4015 4020
Gly Phe Glu Gly Pro Ala Val Ser Val Asp Thr Ala Cys Ser Ser4025 4030 4035Ser Leu Val Ala Leu His Leu Ala Gly Gln Ala Leu Arg Ser Gly4040 4045 4050Glu Cys Ser Met Ala Leu Ala Gly Gly Val Thr Val Met Ser Ser4055 4060 4065Pro Gly Thr Phe Ile Asp Phe Ser Arg Gln Arg Gly Leu Ser Val4070 4075 4080Asp Gly Arg Cys Lys Ser Phe Ala Ala Ala Ala Asp Gly Thr Gly4085 4090 4095Trp Gly Glu Gly Val Gly Met Leu Leu Val Glu Arg Leu Ser Asp4100 4105 4110Ala Glu Arg Asn Gly His Arg Val Leu Ala Val Val Arg Gly Ser4115 4120 4125Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn4130 4135 4140Gly Pro Ser Gln Gln Arg Val Ile Arg Gln Ala Leu Ala Asn Ser4145 4150 4155Gly Leu Thr Gly Ala Asp Val Asp Ala Val Glu Ala His Gly Thr4160 4165 4170Gly Thr Lys Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu Leu Ala4175 4180 4185
Thr Tyr Gly Gln Glu His His Pro Asp Gln Pro Leu Trp Leu Gly4190 4195 4200Ser Leu Lys Ser Asn Ile Gly His Ala Gln Ala Ala Ala Gly Val4205 4210 4215Gly Gly Ile Ile Lys Met Val Met Ala Leu Arg His Glu Thr Leu4220 4225 4230Pro Arg Thr Leu His Ile Asp Glu Pro Thr Pro Gln Val Asp Trp4235 4240 4245Ser Ser Gly Ala Val Ser Leu Leu Thr Glu Pro Arg Pro Trp Pro4250 4255 4260Arg Gln Gly Asp Arg Pro Arg Arg Ala Gly Ile Ser Ser Phe Gly4265 4270 4275Val Ser Gly Thr Asn Ala His Val Ile Leu Glu Glu Ala Pro Ala4280 4285 4290Gln Pro Ala Gly Asp Pro Ala Pro Glu Asp Gly Ala Pro Val Pro4295 4300 4305Trp Ala Met Ser Ala Arg Ser Asn Ala Ala Leu Arg Ala Gln Ala4310 4315 4320Ala Leu Leu Arg Asp Phe Leu Gln Gly Pro Gly Thr Asp Thr Ala4325 4330 4335Leu Arg Ala Val Gly Ala Glu Leu Ala His Gly Arg Ala Val Leu4340 4345 4350
Glu His Arg Ala Val Ile Val Ala Arg Glu Arg Thr Glu Phe Glu4355 4360 4365Asp Ala Leu Glu Ala Leu Ala Ser Gly Glu Pro His Pro Ala Leu4370 4375 4380Ile Glu Asp Thr Thr Gly Ser Gln Thr Asn Ser His Ser Gly Gly4385 4390 4395Gly Val Val Phe Val Phe Pro Gly Gln Gly Gly Gln Trp Ala Gly4400 4405 4410Met Gly Leu Asp Leu Leu Arg Asp Ser Gln Val Phe Ala Asp His4415 4420 4425Val Gly Ala Cys Glu Arg Ala Leu Ala Pro Trp Val Glu Trp Ser4430 4435 4440Leu Thr Glu Met Leu His Arg Asp Ala Glu Asp Pro Val Trp Glu4445 4450 4455Arg Ala Asp Val Val Gln Pro Val Leu Phe Ser Val Met Val Ser4460 4465 4470Leu Ala Ala Leu Trp Arg Ser Tyr Gly Ile Glu Pro Asp Ala Val4475 4480 4485Val Gly His Ser Gln Gly Glu Ile Ala Ala Ala His Val Cys Gly4490 4495 4500Ala Leu Thr Leu Glu Asp Ala Ala Lys Ile Val Ala Leu Arg Ser4505 4510 4515
Arg Ala Leu Ala Ala Leu Arg Gly His Gly Gly Met Ala Ser Leu4520 4525 4530Ala Leu Thr Gly Thr Glu Ala Glu Asp Leu Ile Thr Thr His Trp4535 4540 4545Pro Gly Arg Leu Trp Arg Ala Ala Phe Asn Gly Pro Arg Ala Thr4550 4555 4560Thr Val Ser Gly Asp Thr Asp Ala Leu Asp Glu Leu Leu Thr His4565 4570 4575Cys Thr Glu Thr Gly Val Arg Ala Arg Arg Ile Pro Val Asp Tyr4580 4585 4590Ala Ser His Cys Pro His Thr Glu Thr Ile Glu His Asp Leu Leu4595 4600 4605His Met Leu His Gly Ile Thr Pro Gln Pro Gly Ser Ile Pro Phe4610 4615 4620Tyr Ser Thr Val Glu Asp Ala Trp Thr Asp Thr Thr Thr Leu Asp4625 4630 4635Ala Ala Tyr Trp Tyr Arg Asn Leu Arg Arg Pro Val Arg Phe Thr4640 4645 4650His Ala Val Arg Thr Leu Thr Ala Gln Gly His Arg Leu Phe Ile4655 4660 4665Glu Thr Ser Pro His Pro Thr Leu Thr Pro Ala Ile Glu Asp His4670 4675 4680
Asp His Thr Thr Ala Leu Gly Thr Leu Arg Arg His Asp Asn Asp4685 4690 4695Thr His Arg Phe Leu Thr Ala Leu Ala His Ala His Thr Thr Gly4700 4705 4710His Thr Val Thr Trp Thr Thr His Tyr Pro Thr Thr Pro His Thr4715 4720 4725Pro Ala Ile Asp Leu Pro Thr Tyr Pro Phe Gln His His His Tyr4730 4735 4740Trp Leu His Thr Pro Thr Thr Ser Thr Gly Asp Val Ser Ala Ala4745 4750 4755Gly Leu His Pro Thr Glu His Pro Leu Leu Gly Ala Thr Val Glu4760 4765 4770Leu Ala Asp Gly Asp Gly Thr Leu Leu Thr Gly Arg Leu Ser Leu4775 4780 4785His Thr His Pro Trp Leu Ala Asp His Ser Val Gly Gly Ile Val4790 4795 4800Leu Leu Pro Gly Thr Ala Leu Leu Glu Leu Ala Leu Gln Ala Gly4805 4810 4815Gly Ala Ala His Val Arg Glu Leu Thr Leu His Ala Pro Leu Ala4820 4825 4830Val Pro His Asp Ala Ala Val Asp Leu Gln Val Arg Val Ser Ala4835 4840 4845
Pro Asp Asp Thr Gly Ala Arg Thr Leu Thr Val Ser Ser Arg Ser4850 4855 4860Glu His Ala Arg Pro Glu Asp Pro Trp Gln His His Ala Thr Gly4865 4870 4875Leu Leu Asp Ala Gln Pro Ser Ala Asp Gly Asp Ala Leu Arg Ser4880 4885 4890Trp Pro Pro Glu Gly Ala Leu Pro Cys Ala Ala Asp Glu Leu Glu4895 4900 4905Ser Phe Tyr Ala Ala Gln Glu Ala Arg Gly Phe Ala Tyr Gly Pro4910 4915 4920Ala Phe Arg Gly Leu Arg Ala Ala Trp Arg Arg Gly Glu Glu Val4925 4930 4935Phe Ala Glu Val Arg Leu Pro Glu Ser Val Leu Asp Glu Ala Ser4940 4945 4950Arg Tyr Asn Leu His Pro Ala Leu Leu Asp Ala Ala Leu His Ala4955 4960 4965Val Ala Leu Gly Ala Ala Thr Gly Leu Pro Pro Gly Ala Val Pro4970 4975 4980Phe Ser Phe Ser Gly Val Thr Leu His Ala Val Lys Ala Ala Ala4985 4990 4995Val Arg Val Arg Val Ala Pro Ala Gly Arg Asp Gly Glu Arg Thr5000 5005 50l0
Ala Val Ser Val Ser Leu Ala Asp Glu Thr Gly Arg Gly Val Leu5015 5020 5025Ser Val Asp Ser Leu Ala Val Arg Pro Leu Asp Thr Gly Glu Leu5030 5035 5040Arg Ala Ser Ala Gln Ala Ala Gly Arg Ala Ala Leu Phe Asp Val5045 5050 5055Ala Trp Lys Asp Val Thr Pro Gly Thr Pro Pro Pro Asp Thr Ala5060 5065 5070Val Arg Ser Thr Val Leu Thr His Asp Arg Ala Ala Ala Asp Leu5075 5080 5085Ser Gly Leu Leu Ser Gly Leu Asp Thr Asp Asp Ala Pro Val Pro5090 5095 5100Asp Ala Val Leu Leu Thr Cys Ser Gln Gly Ala Val Ala Asp Val5105 5110 5115Leu Gly Glu Val Leu Ser Val Val Gln Asp Trp Leu Ala Asp Asp5120 5125 5130Arg Leu Ala Glu Ala Arg Leu Val Val Val Thr His Gly Ala Val5135 5140 5145Ala Thr Arg Thr Gly Glu Glu Val Thr Asp Val Ala Gly Ala Ala5150 5155 5160Val Trp Gly Leu Leu Arg Ser Ala Gln Ser Glu His Pro Gly Arg5165 5170 5175
Phe Val Leu Leu Asp Ala Asp Leu Ser Asp Asp Thr Thr Val Thr5180 5185 5190Ala Ala Leu Ala Cys Asp Glu Pro Gln Leu Ala Val Arg Gly Gly5195 5200 5205Arg Leu Leu Ala Ala Arg Leu Ala His Val Pro Val Pro Ala Asp5210 5215 5220Ser Ser Asp Ala Val Arg Phe Asp Ala Glu Gly Thr Val Leu Val5225 5230 5235Thr Gly Gly Thr Gly Thr Leu Gly Ala Ala Val Ala Arg His Leu5240 5245 5250Ala Ala Gly His Gly Val Arg His Leu Leu Leu Val Ser Arg Arg5255 5260 5265Gly Met Ala Ala Thr Gly Ala Glu Glu Leu Cys Ala Glu Leu Gly5270 5275 5280Gln Ala Gly Val Ser Val Ser Val Ala Ala Cys Asp Val Ala Asp5285 5290 5295Arg Ala Gln Val Ala Ala Leu Leu Glu Gln Val Pro Ala Glu His5300 5305 5310Pro Leu Thr Ala Val Val His Thr Ala Gly Val Leu Asp Asp Ala5315 5320 5325Thr Val Ala Cys Leu Asn Arg Glu Lys Ile Asp Ala Val Val Gly5330 5335 5340
Ala Lys Val Asp Gly Ala Leu His Leu His Glu Leu Thr Ala Gly5345 5350 5355Met Asp Leu Ser Ala Phe Val Leu Phe Ser Ser Ala Ala Gly Val5360 5365 5370Leu Gly Ser Pro Gly Gln Gly Asn Tyr Ala Ala Ala Asn Ala Ala5375 5380 5385Leu Asp Ala Leu Ala His Gln Arg Arg Ala Ala Gly Leu Pro Ala5390 5395 5400Leu Ser Leu Ala Trp Gly Leu Trp Glu Glu Ala Ser Gly Met Thr5405 5410 5415Gly His Leu Asp Ala Gly Asp Arg His Arg Ile Thr Arg Ser Gly5420 5425 5430Leu His Pro Leu Thr Thr Pro Asp Ala Leu Ala Leu Leu Asp Thr5435 5440 5445Ala Leu Val Thr Gly Arg Pro Ala Leu Leu Pro Ala Asp Leu Arg5450 5455 5460Pro Thr His Pro Ala Pro Pro Leu Leu Glu His Leu Ala Pro Ala5465 5470 5475Arg Thr Ser Pro Arg Thr Ala His Thr Gly Thr Ser Ala Gly Ala5480 5485 5490Gly Gln Asp Val Ser Leu Ala Asp Arg Leu Ala Thr Leu Thr Pro5495 5500 5505
Glu Gln Gln His Asp Thr Leu Phe Thr Val Val Arg Thr Gln Ile5510 5515 5520Ala Thr Val Leu Gly His Gln Thr Pro Glu Ala Val Pro Ala Asp5525 5530 5535Ser Ala Phe Arg Asp Leu Gly Phe Asp Ser Leu Thr Ala Val Glu5540 5545 5550Leu Arg Asn Arg Leu Ser Arg Ala Thr Gly Leu Arg Leu Pro Ala5555 5560 5565Thr Leu Ala Phe Asp His Pro Thr Ala Thr Ala Leu Thr Arg His5570 5575 5580Leu Leu Thr Arg Leu Leu Pro Asp Asp Ala Ala Thr Ala Pro Pro5585 5590 5595Glu Gln Ser Leu Phe Ala Glu Ile Gly Arg Leu Glu Ala Val Leu5600 5605 5610Ser Ser Val Ala Ser Pro Leu Pro Gly Ala Gln Gly Leu Gly Glu5615 5620 5625Glu Ala Arg Ser Arg Leu Ala Ser Arg Leu Arg Ser Leu Ala Gln5630 5635 5640Val Leu Gly Gly Glu Glu Ala Pro Arg Pro Asp Leu Gly Glu Ala5645 5650 5655Thr Asp Glu Glu Met Phe Ala Leu Ile Asp Gln Glu Thr Gly Ser5660 5665 5670
Pro<210>11<211>5166<212>PRT<213>细菌<400>11Met Ala Asn Glu Glu Met Leu Arg Glu Tyr Leu Lys Arg Ala Thr Ala1 5 10 15Asp Leu Leu Arg Val Arg Arg Arg Leu Glu Gln Val Glu Ser Gly Arg20 25 30Gln Glu Pro Val Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly Gly35 40 45Val Arg Ser Pro Glu Asp Leu Trp Glu Leu Val Ala Ser Gly Gly Asp50 55 60Ala Ile Gly Asp Phe Pro Val Asp Arg Gly Trp Asp Val Glu Asp Leu65 70 75 80Tyr Asp Pro Glu Pro Gly Arg Ala Gly Arg Ser Tyr Thr Arg Ser Gly85 90 95Gly Phe Leu His Glu Ala Ala Glu Phe Asp Ala Gly Phe Phe Gly Leu100 105 110Ser Pro Arg Glu Ala Leu Ala Met Asp Pro Gln Gln Arg Leu Met Leu115 120 125Glu Val Ser Trp Glu Ala Leu Glu Arg Ala Gly Ile Asp Pro Ala Thr
130 135 140Leu Arg Gly Ser Arg Thr Gly Val Phe Ala Gly Met Met Ser His Asp145 150 155 160Tyr Ala Thr Arg Leu Leu Ser Val Pro Asp His Leu Gln Gly Phe Leu165 170 175Gly Asn Gly Asn Ala Ala Ser Val Leu Ser Gly Arg Leu Ser Tyr Thr180 185 190Phe Gly Phe Glu Gly Pro Ala Val Thr Val Asp Thr Ala Cys Ser Ser195 200 205Ser Leu Val Ala Leu His Leu Ala Cys Gln Ser Val Arg Ser Gly Glu210 215 220Ser Ser Leu Ala Leu Ala Gly Gly Val Thr Val Met Ser Thr Pro Ala225 230 235 240Met Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ser Ala Asp Gly Arg245 250 255Cys Lys Pro Tyr Ala Ala Ala Ala Asp Gly Thr Gly Met Ser Glu Gly260 265 270Val Gly Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg Leu Gly275 280 285His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn Gln Asp Gly290 295 300Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser Gln Gln Arg Val
305 310 315 320Ile Gly Gln Ala Leu Val Cys Ala Gly Leu Ser Ala Ala Glu Val Asp325 330 335Val Val Glu Gly His Gly Thr Gly Thr Ser Leu Gly Asp Pro Ile Glu340 345 350Ala Gln Ala Val Leu Ala Ala Tyr Gly Arg Gly Arg Gly Val Pro Leu355 360 365Trp Leu Gly Ser Val Lys Ser Asn Leu Gly His Thr Gln Ala Ala Ala370 375 380Gly Val Ala Gly Val Ile Lys Met Val Met Ala Leu Trp Arg Gly Arg385 390 395 400Leu Pro Arg Thr Leu His Val Asp Glu Pro Ser Pro His Val Asp Trp405 410 415Ser Ser Gly Ala Val Arg Leu Leu Thr Glu Glu Val Val Trp Glu Arg420 425 430Gly Glu Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gly Val Ser Gly435 440 445Thr Asn Ala His Val Ile Leu Glu Glu Ala Pro Gln Glu Glu Glu Val450 455 460Arg Pro Glu Glu Ala Pro Ser Gly Asp Gly Val Gly Pro Val Val Val465 470 475 480Pro Ser Gly Asp Gly Ala Gly Pro Ala Val Val Pro Trp Val Val Ser
485 490 495Ala Arg Ser Glu Ser Ala Leu Arg Gly Gln Ala Arg Arg Leu Arg Val500 505 510Phe Ala Asp Gly Ala Gly Ala Ala Pro Val Glu Val Gly Arg Ala Leu515 520 525Ala Val Glu Arg Ala Trp Leu Glu His Arg Ala Val Val Leu Ala Glu530 535 540Asp Leu Asp Gly Phe Arg His Gly Leu Asp Ala Leu Ala Thr Gly Arg545 550 555 560Pro Ala Pro Glu Val Val Thr Gly Thr Ala Thr Asp Glu Gly Pro Leu565 570 575Ala Phe Leu Phe Ala Gly Gln Gly Thr Gln Arg Pro Ala Met Gly Arg580 585 590Glu Leu His Ala His Phe Pro Ala Phe Ala Asp Ala Phe Asp Glu Val595 600 605Cys Ala His Phe Gly Pro Ile Gly Glu Ala Gly His Thr Leu Arg Asp610 615 620Ile Val Phe Ala Ala Pro Gly Ser Pro Gly Ala Glu Leu Ile Glu Gln625 630 635 640Thr Glu Tyr Ala Gln Pro Ala Leu Phe Ala Val Glu Val Ala Leu Tyr645 650 655Arg Leu Val Glu Asn Trp Gly Val Thr Pro Asp Tyr Leu Leu Gly His
660 665 670Ser Val Gly Glu Leu Ala Ala Ala His Val Ala Gly Met Leu Ser Leu675 680 685Pro Asp Ala Ala Ala Leu Val Thr Ala Arg Gly Arg Leu Met Gln Ala690 695 700Leu Pro Asp Thr Gly Ala Met Val Ala Val Glu Ala Thr Glu Glu Glu705 710 715 720Val Arg Pro Leu Leu Gln Asp Ala Glu Gly Arg Ala Asp Leu Ala Ala725 730 735Val Asn Gly Pro Arg Ala Val Val Leu Ala Gly Asp Glu Asp Ala Val740 745 750Leu Thr Leu Ala Arg His Trp Ala Glu Gln Gly Arg Arg Thr Arg Arg755 760 765Leu Arg Thr Ser His Ala Phe His Ser Pro His Leu Asp Ala Val Leu770 775 780Asp Asp Phe Arg Arg Val Ala Glu Gln Val Val Phe Ala Pro Pro Arg785 790 795 800Ile Pro Val Val Thr Asn Leu Thr Gly Ala Pro Val Ser Ala Asp Thr805 810 815Met Gly Thr Ala Asp Tyr Trp Val Gln His Ala Arg His Thr Val Arg820 825 830Phe Gly Asp Gly Leu Ala Trp Leu Gln Ala Gln Gly Val Thr Ala Tyr
835 840 845Leu Glu Leu Gly Pro Asp Gly Thr Leu Cys Ala Leu Gly Gln Asp Ala850 855 860Leu Thr Glu Pro Ala Pro Leu Leu Pro Ala Leu Arg Pro Asp Arg Pro865 870 875 880Glu Ala Val Ser Val Leu Ala Ala Val Ala Gly Leu Ser Val Arg Gly885 890 895Val Arg Val Asp Trp Ala Ala Val Leu Gly Gly Ala Pro Ser Gly Thr900 905 910Ala Gly Arg Val Glu Leu Pro Thr Tyr Ala Phe Glu Arg Glu Arg Tyr915 920 925Trp Leu Asp Ala Gly Glu Thr Pro Ala Ala Leu Pro Ala Gly Glu Asp930 935 940Gly Pro Leu Trp Gln Ala Val Glu Arg Ala Asp Leu Pro Ala Val Ala945 950 955 960Ala Leu Leu Glu Val Asp Glu Asp Ala Pro Leu Gly Ser Val Val Ser965 970 975Ala Leu Gly Asp Trp Arg Arg Gly Val Arg Glu Arg Ala Val Val Asp980 985 990Gly Trp Arg Tyr Arg Val Val Trp Arg Pro Val Ser Arg Ser Gly Gly995 1000 1005Gly Val Val Ser Gly Gly Val Trp Val Val Val Val Pro Glu Gly
1010 1015 1020Val Val Gly Ala Ala Ala Val Val Glu Gly Leu Glu Arg Ala Gly1025 1030 1035Val Cys Val Arg Val Val Ala Val Glu Gly Gly Cys Ala Asp Arg1040 1045 1050Val Val Leu Gly Glu Arg Leu Arg Glu Val Cys Gly Gly Glu Gly1055 1060 1065Pro Val Gly Val Leu Ala Val Cys Gly Gly Gly Val Gly Val Ala1070 1075 1080Gly Leu Val Leu Gly Leu Val Gln Ala Val Glu Gly Leu Gly Val1085 1090 1095Pro Leu Trp Cys Val Thr Arg Gly Ala Val Ser Val Gly Glu Gly1100 1105 1110Asp Arg Leu Gly Asp Pro Gly Gly Ala Val Val Trp Gly Leu Gly1115 1120 1125Arg Val Ala Gly Leu Glu Leu Pro Asp Arg Trp Gly Gly Val Val1130 1135 1140Asp Leu Pro Glu Val Val Asp Glu Arg Val Val Glu Gly Leu Leu1145 1150 1155Gly Val Leu Ser Gly Gly Gly Gly Glu Gly Glu Val Ala Val Arg1160 1165 1170Ala Ser Gly Val Phe Val Arg Arg Leu Val Arg Ala Pro Gly Gly
1175 1180 1185Gly Ala Glu Ala Gly Gly Trp Arg Pro Arg Gly Thr Val Leu Ile1190 1195 1200Thr Gly Gly Thr Gly Ala Leu Gly Ala His Val Ala Arg Trp Met1205 1210 1215Val Arg Arg Gly Ala Glu His Leu Leu Leu Val Ser Arg Ser Gly1220 1225 1230Arg Glu Ala Lys Gly Ala Gly Glu Leu Arg Ala Glu Leu Thr Ala1235 1240 1245Met Gly Ala Arg Val Thr Ile Ala Ala Cys Asp Val Ala Asp Arg1250 1255 1260Gly Ala Leu Ala Glu Leu Leu Ala Thr Ala Val Pro Glu Asp Cys1265 1270 1275Pro Leu Gly Ala Val Val His Thr Ala Gly Val Val Asp Asp Gly1280 1285 1290Val Leu Asp Ala Leu Thr Pro Glu Arg Leu Glu Gly Val Leu Ala1295 1300 1305Ala Lys Ala Val Gly Ala Arg Asn Leu His Glu Leu Thr Arg Gly1310 1315 1320Ala Asp Leu Ser Ala Phe Val Val Phe Ser Ser Ala Ala Ala Thr1325 1330 1335Phe Gly Ser Gly Gly Gln Gly Ala Tyr Val Ala Ala Asn Ala Tyr
1340 1345 1350Val Glu Ala Leu Ala Val His Arg Arg Gly Leu Gly Leu Pro Ser1355 1360 1365Thr Ala Val Ala Trp Gly Ala Trp Ala Gly Gly Gly Met Ala Ala1370 1375 1380Asp Ala Glu Ala Ala Thr Arg Met Asp Arg Arg Gly Ile Arg Pro1385 1390 1395Met Asp Thr Glu Pro Ala Leu Ser Ala Leu Gly Gln Val Leu Asp1400 1405 1410Arg Asn Glu Thr Cys Leu Thr Ile Ala Asp Ile Asp Trp Glu Arg1415 1420 1425Leu Pro Ala Ala Asp Gly Leu Ala Arg Leu Leu Ser Asp Ile Pro1430 1435 1440Glu Ala Arg Leu Ala Arg Pro Ala Thr Gly Thr Glu Ala Pro Gly1445 1450 1455Ser Leu Arg Ala Arg Leu Ala Ala Leu Glu Pro Ala Glu Arg Asp1460 1465 1470Arg Ala Leu Leu Asp Leu Val Arg Thr His Thr Ala Thr Val Leu1475 1480 1485Gly His Arg Thr Ala Thr Ala Val Pro Ala Asp Arg Ala Phe Arg1490 1495 1500Glu Leu Gly Phe Gly Ser Leu Asn Ala Val Glu Leu Arg Asn Gly
1505 1510 1515Leu Asn Thr Ala Thr Gly Leu Arg Leu Pro Ser Thr Leu Val Phe1520 1525 1530Asp Tyr Pro Asn Pro Ser Ala Leu Ala Thr His Leu Gly Thr Leu1535 1540 1545Leu Ser Thr Gly Gly Glu Ala Pro Ala Gly Arg Pro Ala Phe Ile1550 1555 1560Arg Ser Gly Val Val Asp Glu Pro Val Ala Ile Val Gly Met Ala1565 1570 1575Cys Arg Phe Pro Gly Gly Val Trp Ser Pro Glu Asp Leu Trp Glu1580 1585 1590Leu Val Ala Ser Gly Gly Asp Ala Ile Gly Gly Phe Pro Val Asp1595 1600 1605Arg Gly Trp Asp Val Glu Gly Leu Tyr Asp Pro Glu Ala Gly Arg1610 1615 1620Pro Gly Ser Ser Tyr Thr Arg Ala Gly Gly Phe Leu Ala Gly Ala1625 1630 1635Ala Glu Phe Asp Ala Gly Phe Phe Gly Ile Ser Pro Arg Glu Ala1640 1645 1650Leu Ala Met Asp Pro Gln Gln Arg Leu Leu Leu Glu Val Ser Trp1655 1660 1665Glu Ala Leu Glu Arg Ala Gly Ile Asp Pro Val Ser Leu Arg Gly
1670 1675 1680Ser Arg Thr Gly Val Phe Ala Gly Val Ala Asn Gln Asp Tyr Ala1685 1690 1695Glu Leu Val Arg Arg Gly Gly Arg Asp Leu Glu Gly Tyr Ala Leu1700 1705 1710Thr Gly Val Ser Gly Ser Val Leu Ser Gly Arg Leu Ser Tyr Thr1715 1720 1725Phe Gly Leu Lys Gly Pro Pro Val Thr Val Asn Thr Ala Cys Ser1730 1735 1740Ser Ser Leu Val Ala Leu His Leu Ala Cys Gln Ser Leu Arg Ser1745 1750 1755Gly Glu Ser Lys Leu Ala Leu Pro Gly Gly Val Thr Val Met Ser1760 1765 1770Thr Pro Gly Ala Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ser1775 1780 1785Pro Asp Gly Arg Cys Lys Ala Phe Ala Thr Pro Thr Asn Gly Val1790 1795 1800Gly Trp Ser Glu Gly Val Gly Val Leu Leu Val Glu Arg Leu Ser1805 1810 1815Asp Ala Arg Arg Leu Gly His Arg Val Leu Pro Val Val Arg Gly1820 1825 1830Ser Ala Val Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro
1835 1840 1845Asn Gly Pro Ser Gln Gln Arg Val Ile Gly Gln Ala Leu Val Cys1850 1855 1860Ala Gly Leu Ser Ala Ala Glu Val Asp Val Val Glu Gly His Gly1865 1870 1875Thr Gly Thr Ser Leu Gly Asp Pro Ile Glu Ala Gln Ala Val Leu1880 1885 1890Ala Ala Tyr Gly Arg Gly Arg Gly Val Pro Leu Trp Leu Gly Ser1895 1900 1905Val Lys Ser Asn Leu Gly His Thr Gln Ala Ala Ala Gly Val Ala1910 1915 1920Gly Val Ile Lys Met Val Met Val Leu Trp Arg Gly Arg Leu Pro1925 1930 1935Arg Thr Leu His Val Asp Glu Pro Ser Pro His Val Asp Trp Ser1940 1945 1950Ser Gly Ala Val Arg Leu Leu Thr Glu Glu Val Val Trp Glu Arg1955 1960 1965Gly Glu Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gly Val Ser1970 1975 1980Gly Thr Asn Ala His Val Ile Leu Glu Glu Ala Pro Gln Glu Glu1985 1990 1995Glu Val Arg Pro Glu Glu Ala Pro Ser Gln Gly Glu Ala Gly Pro
2000 2005 2010Ala Val Val Pro Trp Val Val Ser Ala Arg Ser Glu Ser Ala Leu2015 2020 2025Arg Gly Gln Ala Arg Arg Leu Arg Val Phe Ala Asp Gly Ala Gly2030 2035 2040Ala Ala Pro Val Glu Val Gly Arg Ala Leu Ala Val Glu Arg Ala2045 2050 2055Trp Leu Glu His Arg Ala Val Val Leu Ala Glu Asp Leu Asp Gly2060 2065 2070Phe Arg His Gly Leu Asp Ala Leu Ala Thr Gly Leu Pro Thr Ala2075 2080 2085Gly Val Val Ala Gly Arg Thr Gly Pro Glu Ala Asp Gly Lys Ile2090 2095 2100Ala Leu Leu Phe Gly Gly Gln Gly Thr Gln Trp Asp Gly Met Ala2105 2110 2115Ala Glu Leu Leu Asp Ser Ser Pro Val Phe Ala Gln Arg Met Thr2120 2125 2130Glu Cys Ala Asp Ala Leu Arg Pro Tyr Leu Asp Trp Glu Leu Leu2135 2140 2145Asp Val Leu Arg Gly Glu Pro Asp Ala Pro Pro Leu Asp Arg Val2150 2155 2160Asp Val Val Gln Pro Val Leu Phe Ala Val Met Val Ser Leu Ala
2165 2170 2175Ala Leu Trp Arg Ser Tyr Gly Val Arg Pro Asp Ala Val Ala Gly2180 2185 2190His Ser Gln Gly Glu Ile Ala Ala Ala Cys Val Ala Gly Ala Leu2195 2200 2205Ser Leu Glu Asp Ala Ala Arg Val Thr Ala Leu Arg Ser Gln Ala2210 2215 2220Leu Ala Ala Leu Ala Gly Gln Gly Ala Met Ala Ser Val Gly Leu2225 2230 2235Pro Ala Glu Asp Leu Glu Pro Arg Leu Ala Ala Val Asp Pro Ser2240 2245 2250Leu Val Val Ala Ala Asp Asn Gly Ala Arg Ser Ala Val Val Ser2255 2260 2265Gly Ser Pro Asp Ala Val Thr Ala Leu Val Asp Asp Leu Thr Arg2270 2275 2280Asp Gly Val Pro Ala Arg Leu Leu Lys Val Asp Trp Ala Ser His2285 2290 2295Ser Pro Gln Val Glu Ala Ile Arg Ala Asp Leu Leu Gly Leu Leu2300 2305 2310Ala Pro Val Thr Pro Arg Pro Ala Asp Ile Pro Leu Tyr Ser Thr2315 2320 2325Val Thr Gly Glu Pro Val Asp Gly Thr Ala Leu Asp Ala Ala Tyr
2330 2335 2340Trp Tyr Arg Asn Leu Arg Glu Pro Val Arg Phe Arg Asp Ala Thr2345 2350 2355Arg Ala Leu Ala Arg Asp Gly His Thr Val Phe Val Glu Ala Gly2360 2365 2370Pro His Pro Ala Val Ser Val Ala Val Gln Glu Thr Leu Asp Asp2375 2380 2385Leu Gly Ala Ala Asp Thr Leu Val Val Gly Ser Leu Arg Arg Gly2390 2395 2400Glu Gly Gly Leu Arg Arg Phe Leu Ala Ser Ala Ala Glu Leu Ser2405 2410 2415Val Arg Gly Val Arg Val Asp Trp Ala Ala Val Leu Gly Gly Lys2420 2425 2430Pro Ser Gly Thr Ala Gly Arg Val Glu Leu Pro Thr Tyr Ala Phe2435 2440 2445Glu Arg Glu Arg Tyr Trp Leu Asp Pro Glu Glu Thr Pro Ala Ala2450 2455 2460Pro Ala Thr Thr Glu Asp Gly Pro Leu Trp Glu Ala Val Glu Arg2465 2470 2475Glu Asp Pro Ala Ala Val Ala Ala Leu Leu Ala Val Asp Glu Asp2480 2485 2490Ala Pro Leu Asp Ala Leu Val Ser Ala Leu Gly Asp Trp Arg Arg
2495 2500 2505Gly Val Arg Glu Arg Ala Val Val Asp Gly Trp Arg Tyr Arg Val2510 2515 2520Val Trp Arg Pro Val Ser Arg Ser Gly Gly Gly Val Val Ser Gly2525 2530 2535Gly Val Trp Val Val Val Val Pro Glu Gly Val Val Gly Ala Ala2540 2545 2550Ala Val Val Glu Gly Leu Glu Trp Ala Gly Val Cys Val Arg Val2555 2560 2565Val Ala Val Glu Gly Gly Cys Ala Asp Arg Val Val Leu Gly Glu2570 2575 2580Arg Leu Arg Glu Val Trp Gly Gly Glu Gly Pro Val Gly Val Leu2585 2590 2595Ala Val Cys Gly Gly Gly Val Gly Val Ala Gly Leu Val Leu Gly2600 2605 2610Leu Val Gln Ala Val Glu Gly Leu Gly Val Pro Leu Trp Cys Val2615 2620 2625Thr Arg Gly Ala Val Ser Val Gly Glu Gly Asp Arg Leu Gly Asp2630 2635 2640Pro Gly Gly Ala Val Val Trp Gly Leu Gly Arg Val Ala Gly Leu2645 2650 2655Glu Leu Pro Asp Arg Trp Gly Gly Val Val Asp Leu Pro Glu Val
2660 2665 2670Val Asp Glu Arg Val Val Glu Gly Leu Leu Gly Val Leu Ser Gly2675 2680 2685Gly Gly Gly Glu Gly Glu Val Ala Val Arg Ala Ser Gly Val Phe2690 2695 2700Val Arg Arg Leu Val Arg Ala Pro Gly Gly Gly Ala Glu Ala Gly2705 2710 2715Gly Trp Arg Pro Arg Gly Thr Val Leu Ile Thr Gly Glu Asn Ala2720 2725 2730Asp Pro Glu Gln Pro Ala Ala His Leu Ala Arg Trp Leu Ala Asp2735 2740 2745Arg Gly Ala Glu His Leu Leu Leu Ile Ser Thr Ser Gly Asp Gly2750 2755 2760Phe Gly Leu Ala Asp Thr Thr Asp Gln Trp Gly Ala Arg Val Thr2765 2770 2775Ile Ala Ala Cys Asp Val Ala Asp Arg Gly Ala Leu Ala Glu Leu2780 2785 2790Leu Ala Thr Ala Val Pro Glu Asp Cys Pro Leu Gly Ala Val Val2795 2800 2805His Thr Ala Gly Val Val Asp Asp Gly Val Leu Asp Ala Leu Thr2810 2815 2820Pro Glu Arg Leu Glu Gly Val Leu Ala Ala Arg Ala Val Gly Ala
2825 2830 2835Arg Asn Leu His Glu Leu Thr Arg Gly Ala Asp Leu Ser Ala Phe2840 2845 2850Val Val Phe Ser Ser Ala Ala Ala Thr Phe Gly Ser Gly Gly Gln2855 2860 2865Gly Ala Tyr Val Ala Ala Asn Ala Tyr Val Glu Ala Leu Ala Val2870 2875 2880His Arg Arg Gly Leu Gly Leu Pro Ser Thr Ala Val Ala Trp Gly2885 2890 2895Pro Trp Arg Gly His Ser Ala Ala Gly Arg Pro Asp Ala Ala Ala2900 2905 2910Arg Leu His Arg Arg Gly Leu Thr Glu Met Ala Pro Glu Leu Ala2915 2920 2925Leu Ala Ala Leu Ala Arg Val Leu Asp His Asp Glu Ser Gly Leu2930 2935 2940Thr Val Ala Asp Ile Asp Trp Glu Arg Phe Thr Ala His Thr Ala2945 2950 2955Gly Ser Arg Leu Pro Leu Ile Gly Asp Leu Pro Asp Val Arg Ala2960 2965 2970Leu Thr Arg Ala Thr Gly Thr Gly Thr Ala His Gly Thr Asp Leu2975 2980 2985Arg Asp Arg Leu Ala Ala Leu Glu Pro Asp Ala Arg Thr Asp Val
2990 2995 3000Leu Leu Glu Leu Val Ser Thr His Thr Ala Ala Val Leu Gly His3005 3010 3015Arg Glu Ala Asp Thr Val Pro Ala Asp Arg Ala Phe Arg Glu Leu3020 3025 3030Gly Phe Asp Ser Leu Thr Ala Val Glu Leu Arg Asn Arg Leu Asn3035 3040 3045Thr Ala Thr Gly Leu Arg Leu Pro Thr Thr Leu Val Phe Asp Tyr3050 3055 3060Pro Arg Pro Ala Val Leu Ala Arg His Leu Arg Asp Gln Leu Cys3065 3070 3075Gly Thr Ala Pro Ala Thr Pro Pro Val Ala Ala Arg Pro Gly Val3080 3085 3090Val Asp Glu Pro Val Ala Ile Val Gly Met Ala Cys Arg Phe Pro3095 3100 3105Gly Gly Val Trp Ser Pro Glu Asp Leu Trp Glu Leu Val Ala Ser3110 3115 3120Gly Gly Asp Ala Ile Gly Gly Phe Pro Val Asp Arg Gly Trp Asp3125 3130 3135Val Glu Gly Leu Tyr Asp Pro Glu Ala Gly Arg Pro Gly Ser Ser3140 3145 3150Tyr Thr Arg Ser Gly Gly Phe Leu Ala Gly Ala Ala Glu Phe Asp
3155 3160 3165Ala Gly Phe Phe Gly Ile Ser Pro Arg Glu Ala Leu Ala Met Asp3170 3175 3180Pro Gln Gln Arg Leu Leu Leu Glu Val Ser Trp Glu Ala Leu Glu3185 3190 3195Arg Ala Gly Ile Asp Pro Val Ser Leu Arg Gly Ser Arg Thr Gly3200 3205 3210Val Phe Ala Gly Val Ala Asn Gln Asp Tyr Ala Glu Leu Val Arg3215 3220 3225Arg Gly Gly Arg Asp Leu Glu Gly Tyr Ala Leu Thr Gly Val Ser3230 3235 3240Gly Ser Val Leu Ser Gly Arg Leu Ser Tyr Thr Phe Gly Leu Glu3245 3250 3255Gly Pro Ala Val Thr Val Asp Thr Ala Cys Ser Ser Ser Leu Val3260 3265 3270Ala Leu His Leu Ala Cys Gln Ser Leu Arg Ser Gly Glu Ser Glu3275 3280 3285Leu Ala Leu Ala Gly Gly Val Thr Val Met Ser Thr Pro Gly Ala3290 3295 3300Phe Val Glu Phe Ser Arg Gln Arg Gly Leu Ser Ala Asp Gly Arg3305 3310 3315Cys Lys Ala Phe Ala Ala Ala Ala Asp Gly Val Gly Trp Ser Glu
3320 3325 3330Gly Val Gly Val Leu Leu Val Glu Arg Leu Ser Asp Ala Arg Arg3335 3340 3345Leu Gly His Arg Val Leu Ala Val Val Arg Gly Ser Ala Val Asn3350 3355 3360Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ser3365 3370 3375Gln Gln Arg Val Ile Gly Gln Ala Leu Val Cys Ala Gly Leu Ser3380 3385 3390Ala Ala Glu Val Asp Val Val Glu Gly His Gly Thr Gly Thr Ser3395 3400 3405Leu Gly Asp Pro Ile Glu Ala Gln Ala Val Leu Ala Ala Tyr Gly3410 3415 3420Arg Gly Arg Gly Val Pro Leu Trp Leu Gly Ser Val Lys Ser Asn3425 3430 3435Leu Gly His Thr Gln Ala Ala Ala Gly Val Ala Gly Val Ile Lys3440 3445 3450Met Val Met Ala Leu Trp Arg Gly Arg Leu Pro Arg Thr Leu His3455 3460 3465Val Asp Glu Pro Ser Pro His Val Asp Trp Ser Ser Gly Ala Val3470 3475 3480Arg Leu Leu Thr Glu Glu Val Val Trp Glu Arg Gly Glu Arg Pro
3485 3490 3495Arg Arg Ala Gly Val Ser Ser Phe Gly Val Ser Gly Thr Asn Ala3500 3505 3510His Val Ile Leu Glu Glu Ala Pro Gln Glu Glu Glu Val Arg Pro3515 3520 3525Glu Glu Ala Pro Ser Gln Asp Glu Ala Gly Pro Ala Thr Val Pro3530 3535 3540Cys Leu Leu Ser Ala Arg Thr Asp Thr Ala Leu Arg Ala Gln Ala3545 3550 3555Arg Arg Leu Arg Asp Tyr Leu Ala Ala Asn Pro Asp Ile Pro Ile3560 3565 3570Gly Asp Val Ala His Ala Leu Ala Thr Gly Arg Ser Thr Phe Glu3575 3580 3585Arg Arg Ala Val Leu Val Ala Glu Asp His Glu Gly Leu Leu Arg3590 3595 3600Thr Leu Asp Ala Leu Ala Glu Gly Thr Thr Ala Pro Gly Leu Ile3605 3610 3615Glu Ser Pro Ala Arg Thr Ala His Gly Lys Val Ala Phe Leu Phe3620 3625 3630Ser Gly Gln Gly Thr Gln Arg Pro Gly Met Gly Arg Glu Leu Tyr3635 3640 3645Ala Ala His Pro Ala Phe Ala Gln Ala Leu Asp Asp Val Leu Ala
3650 3655 3660Glu Leu Glu Pro His Leu Asp Arg Pro Leu Arg Pro Leu Leu Leu3665 3670 3675Asp Glu Pro Gln Pro Leu Asp Arg Thr Gly Asp Ala Gln Pro Ala3680 3685 3690Leu Phe Ala Leu Gln Val Ala Leu Phe Arg Leu Leu Glu Ser Ala3695 3700 3705Gly Ile Arg Pro Asp His Val Ala Gly His Ser Ile Gly Glu Leu3710 3715 3720Ala Ala Ala His Val Ala Gly Val Leu Ser Leu Thr Asp Ala Ala3725 3730 3735Arg Leu Val Ala Ala Arg Gly Arg Leu Ala Gln Thr Gln Leu Pro3740 3745 3750Pro Gly Gly Ala Met Leu Ala Val Arg Ala Ser Glu Glu Gln Val3755 3760 3765Thr Arg Met Leu Ala Gly Arg Glu Ala Arg Val Ala Val Ala Ala3770 3775 3780Val Asn Gly Pro Thr Ser Val Val Ile Ser Gly Ala Glu Pro Asp3785 3790 3795Val Leu Glu Ala Ala Ala Ala Phe Ala Glu Gln Gly Leu Arg Thr3800 3805 3810Lys Arg Leu Ser Thr Asp Arg Ala Phe His Ser Pro Leu Met Glu
3815 3820 3825Pro Ile Leu Glu Glu Phe Arg Gln Val Ala Thr Gly Ile Ala Tyr3830 3835 3840Ala Glu Pro Thr Ile Pro Val Val Ser Thr Val Thr Gly Asp Arg3845 3850 3855Ala Thr Ala Gly Thr Leu Thr Asp Pro Glu Tyr Trp Val Arg Gln3860 3865 3870Leu Arg Arg Thr Val Arg Phe Gly Asp Ala Val Arg Arg Leu His3875 3880 3885Asp Asp Asp Gly Val Arg Thr Phe Leu Glu Leu Gly Pro Asp Gly3890 3895 3900Thr Leu Cys Ala Leu Ala Gly Glu Cys Leu Pro Ala Asp Asp Asn3905 3910 3915Thr Thr Glu Pro Gly Pro Ala Leu Val Pro Leu Leu Arg Ala Asp3920 3925 3930Arg Pro Glu Pro Leu Ala Leu Leu Thr Ala Leu Ala His Leu His3935 3940 3945Val Gln Gly Thr Pro Lys Gly Gly Thr Ala Val His Trp Pro Ala3950 3955 3960Leu Ile Gly Ala Thr Pro Glu Arg Ala Arg His Leu Asp Leu Pro3965 3970 3975Thr Tyr Pro Phe Asp Arg Arg Arg Tyr Trp Leu Asp Ala Asp Thr
3980 3985 3990Ser Leu Ser Gly Asp Val Ser Ala Ala Gly Leu Thr Ala Ala Gly3995 4000 4005His Pro Leu Leu Gly Ser Ala Val Pro Leu Ala Gly Ser Pro Gln4010 4015 4020Ser Gln Glu Cys Leu Leu Thr Gly Arg Ile Ser Leu Arg Thr His4025 4030 4035Pro Trp Leu Ala Asp His Ala Val Phe Gly Thr Val Leu Leu Pro4040 4045 4050Gly Thr Ala Ile Leu Glu Leu Ala Val Arg Ala Gly Asp Glu Val4055 4060 4065Gly Cys Asp Thr Val Glu Glu Leu Ala Leu Gln Val Pro Leu Val4070 4075 4080Leu Pro Glu Arg Gly Ser Val Val Leu Gln Leu Ser Val Gly Ala4085 4090 4095Thr Glu Thr Ala Pro Asp Gly Val Glu Arg Arg Pro Phe Thr Leu4100 4105 4110Tyr Ala Arg Glu Asp Asp Gly Leu Thr Pro Ala Ala Pro Thr Gly4115 4120 4125Thr Asp Gly Thr Gly Trp Thr Cys His Ala Thr Gly Val Leu Thr4130 4135 4140Arg Arg Ala Glu Thr Ala His Asp Thr Ala Ala Pro Trp Pro Pro
4145 4150 4155Thr Asp Ala Val Pro Val Asp Leu Asp His Trp Tyr Gly Thr Leu4160 4165 4170Ala Asp Ala Gly Leu Gly Tyr Gly Pro Ala Phe Gln Gly Leu Arg4175 4180 4185Ala Ala Trp Arg His Gly Asp Asp Leu Tyr Ala Glu Val Ala Leu4190 4195 4200Pro Asp Gly Pro Ser Gly Asp Ala Asp Arg Tyr Ala Val His Pro4205 4210 4215Ala Leu Leu Asp Ala Ala Leu His Pro Val Val Leu Gly Phe Ala4220 4225 4230Glu Asp Glu Pro Asp Glu Gly His Gly Trp Leu Pro Phe Ser Trp4235 4240 4245Ser Gly Val Thr Val Thr Ala Ser Gly Ala Ser Ala Leu Arg Val4250 4255 4260Arg Leu Ser Arg Arg Ser Pro Asp Thr Ile Ala Leu Leu Ala Thr4265 4270 4275Asp Ser Thr Gly His Thr Val Val Thr Ala Glu Ser Leu Ala Phe4280 4285 4290Arg Pro Val Thr Ala Gly Gln Leu His Ser Ala Arg Thr Ala His4295 4300 4305His Asp Ala Leu Phe Arg Leu Asp Trp Ala Pro Val Pro Leu Pro
4310 4315 4320Arg Thr Pro Ser Ser Lys Thr Arg Leu Ala Leu Ile Gly Ser Glu4325 4330 4335Ala Glu Cys Pro Asp Ala Pro Gly Val Pro Trp Ser Thr Tyr Ala4340 4345 4350Asp Leu Glu Glu Leu Ala Ser Ala Gly Thr Pro Val Pro Asp Val4355 4360 4365Val Val Val Pro Cys Pro His Arg Asp Gly Ala Ala Asp Ala Ala4370 4375 4380Asp Ala Thr Arg Arg Ala Thr Val Arg Val Leu His Leu Leu Gln4385 4390 4395Ser Trp Leu Ala Asp Asp Arg Phe Ala Asp Ser Arg Leu Ala Phe4400 4405 4410Val Thr His Gly Ala Val Ala Ala Ala Pro Gly Asp Ser Val Pro4415 4420 4425Asp Leu Ala His Ala Ala Val Trp Gly Met Val Arg Ser Ala Gln4430 4435 4440Thr Glu Asn Pro Gly Arg Phe Val Leu Thr Asp Leu Asp Asp Thr4445 4450 4455Asp Ala Ser Arg Arg Ala Leu Ala Ala Ala Leu Leu Ser Gly Glu4460 4465 4470Pro Gln Thr Val Leu Arg Glu Gly Arg Ala His Thr Pro Arg Leu
4475 4480 4485Ala Arg Ile Pro Val Gly Ala Arg Ala Asp Ser Gly His Trp Asp4490 4495 4500Pro Asp Ala Thr Val Leu Ile Thr Gly Gly Thr Gly Tyr Leu Gly4505 4510 4515Arg Leu Leu Ala Arg His Leu Val Val Thr His Gly Val Arg His4520 4525 4530Leu Leu Leu Thr Ser Arg Ser Gly Pro Thr Ala Pro Gly Thr Ala4535 4540 4545Glu Leu Val Ala Glu Leu Ala Glu Leu Gly Ala Arg Thr Thr Ala4550 4555 4560Val Ala Cys Asp Leu Ala Asp Arg Arg Ala Val Ala Ala Leu Leu4565 4570 4575Ala Glu Ile Pro Ala Arg His Pro Leu Lys Ala Val Leu His Thr4580 4585 4590Ala Gly Val Val Asp Asp Gly Val Leu Thr Ser Leu Thr Pro Asp4595 4600 4605Arg Leu Asp Ala Val Leu Ser Ala Lys Ala His Gly Ala Ala His4610 4615 4620Leu His Asp Leu Thr Arg Asp Ala Gly Leu Asp Ala Phe Ile Ala4625 4630 4635Phe Ser Ser Ala Ala Ala Ser Phe Gly Ser Pro Gly Gln Ala Asn
4640 4645 4650Tyr Thr Ala Ala Asn Ala Phe Leu Asp Ala Leu Met Gln Gln Arg4655 4660 4665His Ala Leu Gly Leu Pro Gly Arg Ser Leu Ala Trp Gly Arg Trp4670 4675 4680Ala Glu Ala Gly Gly Met Ala Glu His Leu Ala Ala Ala Asp Val4685 4690 4695Ala Arg Met Thr Arg Ser Gly Leu Leu Pro Leu Thr Asn Ala His4700 4705 4710Gly Leu Ala Leu Phe Asp Thr Ala Leu Ala Leu Asp Glu Pro Leu4715 4720 4725Leu Leu Ala Thr Pro Leu Asp Pro Gly Thr Leu Arg Glu Gln Ala4730 4735 4740Ala Val Gly Thr Leu Pro Pro Val Leu Arg Gly Leu Val Arg Thr4745 4750 4755Pro Ala Arg Arg Thr Ala Asp His Gly Val Gly Ala Asp Ala Ala4760 4765 4770Ala Glu Leu Arg Gly Arg Leu Ala Gly Thr Pro Lys Pro Ala Glu4775 4780 4785Arg Thr Ala Leu Leu Thr Glu Val Val Arg Thr His Ala Ala Ala4790 4795 4800Val Leu Gly His Gly Gly Thr Asp Thr Val Thr Ala Asp Gly Glu
4805 4810 4815Phe Arg Glu Phe Gly Phe Asp Ser Leu Thr Ala Val Glu Leu Arg4820 4825 4830Asn Arg Leu Asn Ala Ala Thr Gly Leu Arg Leu Ala Thr Thr Leu4835 4840 4845Val Phe Asp His Pro Thr Pro Ala Ala Leu Ala Asp His Leu Glu4850 4855 4860Arg Leu Leu Ala Ala Glu Pro Ala Ser Asp Met Thr Ala Glu Thr4865 4870 4875Ala Gly Ala Pro Gly Glu Arg Asp Ala Thr Ala Ser Ser Arg Ala4880 4885 4890Gly Ser Gly Pro Ser Ala Asp Thr Val Glu Ala Leu Phe Trp Ile4895 4900 4905Gly His Asp Ser Gly Arg Val Glu Glu Ser Met Ala Leu Leu Ser4910 4915 4920Ala Ala Ser Ala Phe Arg Pro Cys Phe Thr Asp Pro Ser Ala Met4925 4930 4935Thr Arg Pro Pro Phe Val Arg Val Ala Gln Gly Asp Thr Gly Pro4940 4945 4950Ala Leu Ile Cys Leu Pro Thr Val Ala Ala Val Ser Ser Val Tyr4955 4960 4965Gln Tyr Ser Arg Phe Ala Ala Ala Leu Asp Gly Leu Arg Asp Val
4970 4975 4980Trp Tyr Val Pro Ala Pro Gly Phe Ala Asp Gly Glu Pro Leu Pro4985 4990 4995Ala Asp Val Asp Thr Ile Thr Arg Leu Phe Thr Asp Ala Ile Leu5000 5005 5010Arg His Thr Asp Gly Glu Pro Phe Ala Leu Ala Gly His Ser Ala5015 5020 5025Gly Gly Trp Phe Thr His Thr Val Thr Ser Arg Leu Glu His Leu5030 5035 5040Gly Val Arg Pro Gln Ala Val Val Val Met Asp Ala Tyr Leu Pro5045 5050 5055Asp Glu Gly Met Ala Pro Val Ala Ala Ala Leu Thr Ser Glu Ile5060 5065 5070Phe Asp Arg Val Thr Glu Phe Ile Asp Leu Asp Tyr Ala Arg Leu5075 5080 5085Val Ala Met Gly Gly Tyr Phe Arg Ile Phe Ala Gly Trp Arg Pro5090 5095 5100Pro Ala Leu Glu Thr Pro Thr Leu Phe Leu Arg Ala Arg Glu Ser5105 5110 5115Glu Gln Pro Pro Pro Val Trp Gly Glu Pro His Thr Val Leu Glu5120 5125 5130Thr Asp Gly Asn His Phe Thr Met Leu Glu Glu His Ala Glu Ser
5135 5140 5145Thr Ala Arg His Val His Thr Trp Leu Ala Gly Leu Thr Glu Gln5150 5155 5160Arg Arg Arg5165<210>12<211>254<212>PRT<213>细菌<400>12Met Asp Arg Tyr Ala Lys Arg Phe Glu Asp Arg Leu Val Leu Val Thr1 5 10 15Gly Ala Gly Ser Gly Ile Gly Arg Ala Thr Ala Cys Arg Phe Gly Ala20 25 30Ala Gly Ala Arg Leu Val Cys Val Asp Arg Asp Gly Pro Gly Ala Glu35 40 45Ala Thr Ala Glu Leu Ala Arg Ala Arg Gly Ala Arg Ala Ala Cys Ala50 55 60Glu Val Ala Asp Val Ser Asp Glu Val Ala Met Glu Arg Leu Ala Ala65 70 75 80Arg Val Thr Ala Ala His Gly Val Leu Asp Val Leu Val Asn Asn Ala85 90 95Gly Ile Gly Met Ser Gly Arg Phe Leu Asp Thr Ser Ala Glu Asp Trp100 105 110
Arg Arg Thr Leu Gly Val Asn Leu Trp Gly Val Ile His Gly Cys Arg115 120 125Leu Leu Gly Arg Gly Met Ala Glu Arg Arg Gln Gly Gly His Ile Val130 135 140Thr Val Ala Ser Ala Ala Ala Phe Gln Pro Thr Arg Val Val Pro Val145 150 155 160Tyr Ala Thr Ser Lys Ala Ala Ala Leu Met Leu Ser Glu Cys Leu Arg165 170 175Ala Glu Leu Ala Glu Phe Gly Ile Gly Val Ser Val Val Cys Pro Gly180 185 190Leu Val Arg Thr Pro Phe Ala Ser Ala Met Tyr Phe Ala Gly Ala Ser195 200 205Pro Asp Glu His Thr Arg Leu Arg Glu Ser Ser Ala Arg Arg Phe Ala210 215 220Gly Arg Gly Cys Pro Pro Glu Lys Val Ala Asp Ala Val Leu Arg Ala225 230 235 240Ile Met Arg Thr Ala Leu Pro Thr Val Thr Gly Ser Thr Pro245 250<210>13<211>7<212>PRT<213>细菌<400>13
Gly Gly Thr Gly Thr Leu Gly1 5<210>14<211>7<212>PRT<213>细菌<400>14Gly Ala Ala Ser Thr Leu Gly1 5<210>15<211>33<212>DNA<213>细菌<400>15ctggtgacgg gcgctgcaag cactctgggg gcg 33<210>16<211>33<212>DNA<213>细菌<400>16gaccactgcc cgcgacgttc gtgagacccc cgc 33<210>17<211>7<212>PRT<213>细菌<400>17Leu Val Ser Arg Arg Gly Met1 5
<210>18<211>7<212>PRT<213>细菌<400>18Leu Val Ala Ala Ala Gly Met1 5<210>19<211>47<212>DNA<213>细菌<400>19gcggcatctg ctgctggtgg cagcggcagg catggccgcc gccggtg 47<210>20<211>47<212>DNA<213>细菌<400>20cgccgtagac gacgaccacc gtcgccgtcc gtaccggcgg cggccac 47<210>21<211>7<212>PRT<213>细菌<400>21His Thr Ala Gly Val Leu Asp1 5<210>22<211>7<212>PRT
<213>细菌<400>22His Thr Pro Pro Leu Leu Asp1 5<210>23<211>46<212>DNA<213>细菌<400>23gaccgctgtg gtgcacacgc cacctctcct ggacgacgcc accgtg46<210>24<211>46<212>DNA<213>细菌<400>24ctggcgacac cacgtgtgcg gtggagagga cctgctgcgg tggcac46<210>25<211>5<212>PRT<213>细菌<400>25Gly Ala Lys Val Asp1 5<210>26<211>5<212>PRT<213>细菌<400>26
Gly Ala Ala Val Asp1 5<210>27<211>39<212>DNA<213>细菌<400>27gatgcggtgc tcggggcggc tgtggacggt gccctgcac39<210>28<211>39<212>DNA<213>细菌<400>28ctacgccacg agccccgccg acacctgcca cgggacgtg39<210>29<211>7<212>PRT<213>细菌<400>29Val Leu Phe Ser Ser Ala Ala1 5<210>30<211>7<212>PRT<213>细菌<400>30Val Leu Phe Ala Ala Ala Ala1 5
<210>31<211>41<212>DNA<213>细菌<400>31gtcggcgttc gtgctgttcg cagcggccgc cggggtcctg g 41<210>32<211>41<212>DNA<213>细菌<400>32cagccgcaag cacgacaagc gtcgccggcg gccccaggac c 41<210>33<211>23<212>DNA<213>细菌<400>33tactgcgcca cacggagccc gag 23<210>34<211>20<212>DNA<213>细菌<400>34tgggtaacgc cagggttttc 20<210>35<211>24<212>DNA<213>细菌<400>35ggaaacagct atgacatgat tacg24
<210>36<211>20<212>DNA<213>细菌<400>36tcggagccgc tccacctgag 20<210>37<211>20<212>DNA<213>细菌<400>37cctgatggac gcgggtgcgc 20<210>38<211>16<212>DNA<213>细菌<400>38gacaccgaaa cccctg 16<210>39<211>20<212>DNA<213>细菌<400>39cctgatggac gcgggtgcgc 20<210>40<211>23<212>DNA<213>细菌<400>40gccgtgtgca ccacagcggt cag 23
<210>41<211>28<212>DNA<213>细菌<400>41gtgtgatgtc gccgaccgcg cccaggtc28<210>42<211>22<212>DNA<213>细菌<400>42gcgctggtgg gccagggcgt cc 2
权利要求
1.编码用于生产LL-F28249化合物的生物合成途径中的至少一种蛋白质的经纯化和分离的核酸分子，其中所述的核酸分子是从生产抗生素的野生型或突变的链霉菌中分离出来的。
2.根据权利要求1所述的核酸分子，其中所述的核酸分子是从生产抗生素的野生型或突变的蓝灰链霉菌非产蓝亚种中分离出来的。
3.根据权利要求1所述的核酸分子，其中所述的LL-F28249化合物是LL-F28249α。
4.根据权利要求1所述的核酸分子，其中所述的分子具有SEQ ID NO1所示序列或其互补序列。
5.与权利要求4的核酸分子的序列杂交的并且编码用于生产LL-F28249化合物的生物合成途径中的一种蛋白质的核酸分子。
6.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的7697-10465的核苷酸。
7.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的10791-11570的核苷酸。
8.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的11659-12462的核苷酸。
9.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的12850-19875的核苷酸。
10.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的19876-31036的核苷酸。
11.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的31115-49246的核苷酸。
12.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的50449-51303的核苷酸。
13.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的51300-52706的核苷酸。
14.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的52809-69833的核苷酸。
15.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的69929-85429的核苷酸。
16.权利要求4所述的核酸分子，其中所述的分子包括SEQ ID NO1的85574-86338的核苷酸。
17.含有权利要求1所述的核酸分子的生物功能质粒或载体。
18.根据权利要求17所述的质粒或载体，其中所述的质粒或载体包括具有ATCC指定号为PTA-4392的Cos11、具有ATCC指定号为PTA-4393的Cos36或具有ATCC指定号为PTA-4394的Cos40。
19.被权利要求17所述的质粒或载体稳定转化或转染的适当的宿主细胞。
20.根据权利要求19所述的宿主细胞，其中所述的宿主是埃希氏菌属、放线菌、芽孢杆菌属、棒状菌或嗜热放线菌属。
21.根据权利要求20所述的宿主细胞，其中所述的宿主是大肠杆菌、浅青紫链霉菌、天蓝色链霉菌、灰褐链霉菌及产二素链霉菌。
22.由权利要求1所述的核酸分子编码的生物合成蛋白质。
23.根据权利要求22所述的蛋白质，其中开放阅读框的氨基酸序列如SEQID NO2至SEQ ID NO12中的任意一个所示，或是其具有生物活性的变异体。
24.一种LL-F28249化合物生物合成所涉及蛋白质的生产方法，所述方法包括在适合的营养条件下，以一种允许蛋白质产物表达的方式，培养由权利要求1所述的核酸分子转化或转染的原核或真核宿主细胞，然后分离该核酸分子表达的目的蛋白产物。
25.权利要求24所述的原核或真核宿主细胞中的核酸分子表达的蛋白质产物。
26.用于克隆编码LL-F28249化合物的生物合成途径中的蛋白质的核酸分子的一种质粒或者是两种或三种质粒的组合，其中所述质粒或组合包含跨越整个生物合成基因簇并编码用于生产LL-F28249化合物的I型聚酮合酶的核酸分子。
27.根据权利要求26所述的组合，其包含具有ATCC指定号为PTA-4392的Cos11，具有ATCC指定号为PTA-4393的Cos36以及具有ATCC指定号为PTA-4394的Cos40。
28.一种用以制备23-氧-LL-F28249化合物的方法，其包括以下步骤(a)对权利要求1所述的核酸分子的组件3酮还原酶酶域进行突变，以构建使23-酮还原酶域无功能的突变的核酸分子；(b)用突变的核酸分子转化或转染生产抗生素的野生或突变的链霉菌菌株，从而用突变的组件3酮还原酶域替换原始的组件3酮还原酶域；(c)以一种能允许产生23-氧-LL-F28249化合物的方式，在适合的营养培养基中培养经转化或转染的链霉菌菌株；以及(d)回收23-氧-LL-F28249化合物。
29.根据权利要求28的方法，其中所述的23-氧-LL-F28249化合物是23-氧-LL-F28249α。
30.一种用于制备23-氧-LL-F28249化合物的方法，其包括以下步骤(a)对权利要求1所述的核酸分子的组件3酮还原酶域进行突变，以构建使23-酮还原酶域无功能的突变的核酸分子；(b)用突变的核酸分子转化或转染合适的原核或真核宿主细胞；(c)以一种允许23-氧-LL-F28249化合物表达的方式，在适合的营养培养基中培养经转化或转染的宿主细胞；以及(d)回收23-氧-LL-F28249化合物。
31.根据权利要求30的方法，其中所述的23-氧-LL-F28249化合物是23-氧-LL-F28249α。
32.一种用于制备23-(O-甲基肟)-LL-F28249化合物的方法，其包括以下步骤(a)对权利要求1所述的核酸分子的组件3酮还原酶域进行突变，以构建使23-酮还原酶域无功能的突变的核酸分子；(b)采用突变的核酸分子转化或转染生产抗生素的野生或突变的链霉菌菌株，从而用突变的组件3酮还原酶域替换原始的组件3酮还原酶域；(c)以一种能允许产生23-氧-LL-F28249化合物的方式，在适合的营养培养基中培养经转化或转染的链霉菌菌株；(d)回收23-氧-LL-F28249化合物；(e)在适当的反应条件下，将23-氧-LL-F28249化合物转变成23-(O-甲基肟)-LL-F28249化合物；以及(f)分离23-(O-甲基肟)-LL-F28249化合物。
33.根据权利要求32所述的方法，其中所述的23-氧-LL-F28249化合物是23-氧-LL-F28249α并且所述的方法用来制备23-(O-甲基肟)-LL-F28249α化合物。
34.一种用于制备23-(O-甲基肟)-LL-F28249化合物的方法，其包括以下步骤(a)对权利要求1所述的核酸分子的组件3酮还原酶酶域进行突变，以构建使23-酮还原酶域无功能的突变的核酸分子；(b)采用突变的核酸分子转化或转染合适的原核或真核宿主细胞；(c)以一种能允许表达23-氧-LL-F28249化合物的方式，在适合的营养培养基中培养经转化或转染的宿主细胞；(d)回收23-氧-LL-F28249化合物；(e)在适当的反应条件下，将23-氧-LL-F28249化合物转变成23-(O-甲基肟)-LL-F28249化合物；以及(f)分离该23-(O-甲基肟)-LL-F28249化合物。
35.根据权利要求34所述的方法，其中所述的23-氧-LL-F28249化合物是23-氧-LL-F28249α并且所述的方法用来制备23-(O-甲基肟)-LL-F28249α化合物。
全文摘要
本发明涉及用以形成LL－F28249化合物以及最重要的其主要成分LL－F28249α的完整的生物合成途径。经纯化和分离的，编码该生物合成途径蛋白的核酸分子在SEQ ID NO1中被完整的描述，该核酸分子是分离自野生型或突变链霉菌属(Streptomyces)。该DNA基因簇及其在合适宿主中的表达，使得高活性的天然代谢物以及半合成衍生物的有效生产成为可能。本发明进一步涉及那些含有以及表达这种新的核酸分子的质粒、载体和宿主细胞。尤其感兴趣的是，整个生物合成途径紧密地装配在Cos11，Cos36和Cos40这三个质粒中。本发明还涉及那些经纯化和分离的由该完整的DNA基因簇编码的经纯化和分离的生物合成蛋白质。此外，本发明涉及一种新的，有效制备莫西菌素的生物化学方法。
文档编号A61P31/04GK1676607SQ20041004779
公开日2005年10月5日 申请日期2004年5月14日 优先权日2003年5月16日
发明者C·黄, D·T·查勒夫, M·E·鲁彭, J·斯蒂芬斯 申请人:惠氏公司