本发明涉及生物技术领域,尤其是涉及一种原发性特应性疾病相关基因的捕获探针、试剂盒及其应用。
背景技术:
原发性特应性疾病(primaryatopicdisorders,pads)是单个基因变异所致的遗传性疾病。除了通常认为的免疫耐受机制缺陷外,参与调节皮肤屏障、肥大细胞功能、tcr信号及细胞骨架、细胞因子信号通路的基因缺陷,都可能表现为pads。不过,由于基因的外显率不同,基因、环境间相互作用下可出现不完全相同的临床表型。pads相关37个基因主要包括flg、cdsn、dsg1、dsp、spink5、plcg2、adgre2、tpsab1、foxp3、il2ra、tgfbr1、tgfbr2、was、malt1、card11、wipf1、arpc1b、dock8、carmil2、erbin、znf341、rag1、rag2、dclre1c、ada、il7ra、chd7、lig4、zap70、stat3、statl、stat5b、jak1、il4ra、il6st、il6r、pgm3。
flg基因编码丝聚蛋白(filaggrin,flg)。flg蛋白由角质形成细胞合成,flg蛋白可以分解产生大量天然保湿因子参与皮肤屏障,flg基因突变致flg蛋白表达降低也可以引起特异性皮炎、寻常性鱼鳞病等。
cdsn基因、dsp基因和dsg1基因编码三种桥粒蛋白,皮肤通过桥粒蛋白构成的桥粒锚定来抵抗机械应力。这些基因突变可引起严重的特应性皮炎、ige升高和多种过敏。
spink5基因编码一种丝氨酸蛋白酶抑制剂lektl,lekt1靶向作用于激肽释放酶5(kallikreins5,klk5)。激肽释放酶是最重要的表皮细胞蛋白酶之一,蛋白酶及其抑制分子维持表皮的脱屑过程,在皮肤屏障中发挥重要作用。纯合的spink5基因变异使表皮蛋白酶活性失调而致nethron综合征,患者表现出过敏(严重的过敏性皮炎、花粉过敏、食物过敏等),伴随先天性鱼鳞病红皮病、竹节样毛发。
plcg2基因编码磷脂酶cγ2(plcγ2)——一种在b细胞、自然杀伤细胞及肥大细胞中表达的信号分子。plcg2基因缺失通过暴露磷脂酶c(plc)磷酸化位点,引发寒冷相关的荨麻疹,可伴哮喘、过敏性鼻炎等表现及肥大细胞增多、ige升高。
adgre2基因编码粘附g蛋白偶联受体e2基因,该基因错义突变表现为运动或擦毛巾等诱发局部红斑、瘙痒性荨麻疹。
tpsab1基因编码α胰蛋白酶,该基因拷贝数增加引起遗传性高类胰蛋白酶血症,表现为类胰蛋白酶水平升高、肥大细胞活性增加,在振动刺激下出现荨麻疹。
foxp3基因编码转录因子foxp3,foxp3是控制treg细胞发育和功能的关键转录因子之一。foxp3基因变异致treg细胞减少或缺乏,引起x连锁隐性遗传的多内分泌腺病肠病伴免疫失调综合征(immunodysregμlation,polyendocrinopathy,entropathy,x-1inkedsyndrome,ipex)。ipex除自身免疫疾病表现外,尚伴严重特应性皮炎、高ige和嗜酸性粒细胞增多。
il2ra基因编码il-2受体α链(il2ra,cd25),treg细胞的cd25结合il-2使效应t细胞il-2r缺乏配体而凋亡,故il2ra基因及stat5b基因的功能缺失性突变使treg抑制作用减弱,与ipex类似,表现出免疫失调及特应性疾病。
stat1基因、stat3基因、stat5b基因分别编码信号传导与转录活化因子(stat)蛋白家族的成员。stat1gof抑制th17细胞,引起慢性皮肤粘膜念珠菌病(chronicmucocutaneouscandidiasis,cmc),特征为引起指甲、皮肤、口腔和生殖器粘膜反复或持续念珠菌感染。stat3基因突变是常染色体显性遗传高ige综合征(hies)的主要原因。stat3基因缺失性变异使其磷酸化及转录受抑,il-4分泌增加,ige大量分泌,th17分泌减少,临床表现为慢性湿疹样皮炎及复发性葡萄球菌样皮肤脓肿、复发性细菌性肺部感染,伴结缔组织异常,包括头面部畸形、关节过度活动、动脉瘤形成等。stat5b变异致il-2基因转录受抑,可致身材矮小、严重的免疫失调。
tgfbr1基因和tgfbr2基因编码tgfβ受体,其基因突变引起常染色体显性遗传的loeys-dietz综合征,表现为主动脉瘤和全身性动脉迂曲、超长,悬雍垂裂或腭裂、伴有反复呼吸道感染,湿疹,食物过敏。
肌动蛋白细胞骨架相关基因(dock8、was、carmil2、wipf1、arpc1b)缺陷致免疫突触形成受阻从而抑制t细胞信号传导。dock8基因突变致常染色体隐性遗传的高ige综合征(hyperigesyndromes,hies),临床表现为反复感染、过敏性疾病、湿疹和食物过敏、嗜酸性粒细胞增多症。was基因突变致wiskott-aldrich综合征,wipf1基因编码的蛋白与wiskott-aldrich综合征中经常突变的wiskott-aldrich综合征蛋白区域结合,这两种蛋白质之间相互作用的削弱可能导致该疾病,表现为反复严重湿疹,arpc1b基因纯合突变致was相似的表型。支架蛋白carmil2缺陷表现为特应性皮炎、过敏性哮喘、食物过敏等表型。除pads外,上述基因缺陷致肌动蛋白合成障碍,t淋巴细胞分化增殖能力下降,亦伴有联合免疫缺陷和自身免疫的症状。
card11基因编码card11蛋白,malt1编码malt1蛋白。card11蛋白和malt1蛋白是t细胞受体信号通路中的重要分子,card11基因和malt1基因突变阻碍cbm(card11-bcl10-malt1)复合物形成,nf-κb激活受阻,t细胞活化受到影响。card11基因和malt1基因突变的患儿存在严重的特应性疾病,包括变应性鼻炎、食物过敏、特应性皮炎、嗜酸性食管炎等,由于t细胞功能受损,患者常合并联合免疫缺陷,表现为反复细菌和念珠菌感染(肺部、皮肤和胃肠道)、牙周疾病(口疮、唇炎、牙龈炎)和发育不良。
il6r基因编码il-6rα,il6st基因编码跨膜受体gp130,gp130与il-6rα共同构成il-6r,il-6r与il6st基因的缺失型变异使stat3磷酸化受阻致hies,亦表现为反复细菌感染、结缔组织异常及ige升高。
il4ra基因编码白细胞介素4(il-4)的受体,该基因功能获得性变异致th2分化,产生ige,可表现为一系列过敏性疾病。
jak1基因编码人类酪氨酸激酶蛋白(jak1),jak1基因的功能获得性变异促进il-4激活、ige升高,表现为明显的嗜酸性粒细胞增多、严重的特应性皮炎及生长发育障碍。
erbin基因编码erbb2相互作用蛋白,富含亮氨酸重复序列和pdz结构域(lap)家族的成员,编码的蛋白质含有17个富含亮氨酸的重复序列和一个pdz结构域。它与erbb2蛋白的非磷酸化形式结合并调节erbb2的功能和定位。它也被证明通过破坏ras-raf相互作用影响ras信号通路。
znf341基因是细胞核内的转录因子,激活stat3的启动子,znf341变异使stat3不被正常激活致stat3dn类似的hies,表现为ige显著升高,反复感染及结缔组织异常,但结缔组织异常不如stat3变异显著。
rag1和rag2是tcr重排所必需的,rag1、rag2基因突变使其编码的重组酶活性完全或部分丧失,vdj重排失衡。ragl、rag2基因突变致常染色体隐性遗传的omenn综合征,表现为t-b-nk+重症联合免疫缺陷,伴有湿疹、红皮病、脱发、肝脾肿大等表现,可见嗜酸性粒细胞增多和ige水平升高。
腺苷酸脱氨酶(adenosinedeaminase,ada)缺陷致三磷酸脱氧腺苷酸堆积从而抑制dna合成必需的核糖核苷还原酶,引起常染色体隐性遗传的t-b-nk-重症联合免疫缺陷,表现为反复机会性感染、念珠菌病及淋巴组织缺如或发育不全,伴哮喘、鼻窦炎及ige升高,部分可有骨骼异常、肺泡蛋白沉积症等。
zap70基因突变使zap70蛋白表达缺失,tcr信号转导障碍,表现为联合免疫缺陷、反复感染、腹泻及肝脾肿大,伴有皮肤红斑样及湿疹样改变。
il7ra基因编码il-7受体,il-7促进胸腺细胞成熟,il7ra基因突变使il-7受体不能转导刺激胸腺细胞成熟的信号致t细胞发育成熟障碍,致t-b+nk+的联合免疫缺陷,可致中耳炎、鹅口疮、腹泻、反复感染及发生机会性感染,伴皮肤、头发、指甲等疱疹样皮炎等改变。
chd7基因染色质解旋酶dna结合蛋白7,该基因编码包含几个解旋酶家族结构域的蛋白质,目前已有两个转录本,在一些charge综合征患者中发现了该基因的突变。
artemis-dna-pk参与t、b细胞分化,与之相关的基因缺陷有lig4基因和dclre1c基因,使t细胞、b细胞分化停滞,致t-b-nk+重症联合免疫缺陷。dclre1c基因编码artermis致omenn综合征,lig4基因编码dna连接酶ⅳ,存在小头畸形及银屑病、毛细血管扩张等皮肤表现。
磷酸谷胱甘肽酶(pgm3)是氨基己糖生物合成的关键下游介质,对尿苷二磷酸(udp)n-乙酰氨基葡萄糖(glcnac)的生成至关重要。甘露糖蛋白n-乙酰氨基葡萄糖转移酶(mgat)与udp-glcnac结合为n-聚糖,限制t细胞激活和glut1的表达。pgm3lof致udp-g1cnac及n-聚糖减少,促进th2分化,引起特应性疾病(湿疹、哮喘、多系统过敏等)及ige升高,部分可见骨骼异常(脊柱侧弯、短指等),常伴随神经系统表现(共济失调、肌张力减退、构音障碍等)。
技术实现要素:
本发明提供了一种原发性特应性疾病相关基因的捕获探针,所述原发性特应性疾病相关基因选自flg、cdsn、dsg1、dsp、spink5、plcg2、adgre2、tpsab1、foxp3、il2ra、tgfbr1、tgfbr2、was、malt1、card11、wipf1、arpc1b、dock8、carmil2、erbin、znf341、rag1、rag2、dclre1c、ada、il7ra、chd7、lig4、zap70、stat3、stat1、stat5b、jak1、il4ra、il6st、il6r、pgm3中的两种或多种。
可选地,根据上述的捕获探针,所述捕获探针为如下1)或2);
1)由序列表中seqidno.1至seqidno.394所示的探针中的两种或多种组成;
2)由1)中每条探针的衍生物中的两种或多种组成;
所述每条探针的衍生物为将1)中每条探针经过一个或几个核苷酸的取代和/或缺失和/或添加且具有相同功能的探针。
所述几个核苷酸的取代和/或缺失和/或添加可为二至十个核苷酸的取代和/或缺失和/或添加,例如可为二、三、四、五、六、七、八、九或十个核苷酸的取代和/或缺失和/或添加。
上述捕获探针中,所述每条探针均可标记生物素。
本发明还提供了一种原发性特应性疾病相关基因捕获试剂盒,包括上述的捕获探针。
可选地,根据上述的试剂盒,所述试剂盒还包括如下独立包装的各个组分中至少一种:
2)富集缓冲液bl,其包括人cot-1dna、鲑鱼精dna、用于封闭接头序列的引物;
3)富集缓冲液hy,包括nacl、柠檬酸钠、bsa和tween20;
4)文库富集结合缓冲液,包括nacl、edta和tris-hcl缓冲液;
5)洗涤缓冲液wb1,包括sds和柠檬酸三钠缓冲液;
6)洗涤缓冲液wb2,包括sds和10倍稀释的所述柠檬酸三钠缓冲液;
7)洗涤缓冲液wb3,包括sds和2倍稀释的所述柠檬酸三钠缓冲液;
8)文库富集洗脱液,其为naoh水溶液;
9)中和缓冲液ne,其为tris-hcl缓冲液;
10)pcr反应液,包括用于pcr扩增的引物。
可选地,上述富集缓冲液bl中用于封闭接头序列的引物为引物1和引物2,引物1的序列为seqidno.397,引物2的序列为seqidno.398。
可选地,上述富集缓冲液bl由30%-45%(v/v)(如30%-35%、35%-45%、30%、35%、40%、45%)人cot-1dna、5%-25%(v/v)(如5%-15%、15%-25%、5%、10%、15%、20%、25%)鲑鱼精dna、0.2-1nmol/μl(如0.2nmol/μl、0.3nmol/μl、0.5nmol/μl、0.6nmol/μl、1nmol/μl)引物1、0.2-1nmol/μl(如0.2nmol/μl、0.3nmol/μl、0.5nmol/μl、0.6nmol/μl、1nmol/μl)引物2和水组成。
可选地,上述富集缓冲液hy由1-1.5m(如1.0-1.25m、1m、1.25m、1.5m)nacl、0.1-0.3m(如0.1-0.15m、0.15-0.3m、0.1m、0.125m、0.15m、0.3m)柠檬酸钠、0.08-0.12g/100ml(如0.08、0.1、0.12)bsa、5%-9%(v/v)(如5-7%、7-9%、5%、6%、7%、8%、9%)tween20和水组成。
可选地,上述文库富集结合缓冲液由1mnacl、1mmedta和ph7.5、10mm的tris-hcl缓冲液组成。
可选地,上述洗涤缓冲液wb1为质量体积比(质量体积比g:ml)0.05-0.2%的sds溶液a(如0.05-0.1%、0.1-0.2%、0.05%、0.1%、0.2%);0.05-0.2%的sds溶液a中的溶质为sds,浓度为0.05-0.2%(如0.05-0.1%、0.1-0.2%、0.05%、0.1%、0.2%),溶剂为柠檬酸三钠缓冲液ssc。
可选地,上述洗涤缓冲液wb2为质量体积比0.05-0.2%sds溶液b;sds溶液b中的溶质为sds,浓度为0.05-0.2%(如0.05%、0.06%、0.1%、0.15%、0.2%),溶剂为0.1×ssc。
可选地,上述洗涤缓冲液wb3为质量体积比0.05-2%(如0.05%,0.08,0.1%,0.15%,0.2%)sds溶液c;sds溶液c中的溶质为sds,浓度为0.05-2%,溶剂为0.5×ssc。
可选地,文库富集洗脱液为0.1mnaoh水溶液。
可选地,中和缓冲液ne为ph值为7.5且浓度为1mtris-hcl水溶液。
可选地,pcr反应液中的用于pcr扩增的引物为引物3和引物4,引物3的序列为seqidno.399,引物4的序列为seqidno.400,且所述引物3和所述引物4最后一位核苷酸均为硫代修饰。
上述的捕获探针、上述的试剂盒在制备捕获待测样本中原发性特应性疾病相关基因的产品中的应用是本发明保护的范围。
上述的捕获探针、上述的试剂盒在制备测序待测样本中原发性特应性疾病相关基因产品中的应用也是本发明保护的范围。
上述的捕获探针、上述的试剂盒在制备检测或预测待测者是否患有原发性特应性疾病产品中的应用也是本发明保护的范围。
本发明还提供了一种捕获待测样本中原发性特应性疾病相关基因的方法,包括如下步骤:
利用上述的捕获探针对待测样本的基因组dna进行捕获,得到捕获片段;实现捕获待测样本中原发性特应性疾病相关基因。
本发明还提供了一种测序待测样本中原发性特应性疾病相关基因或基因突变位点的方法,包括如下步骤:
1)利用上述的捕获探针对待测样本的基因组dna进行捕获,得到捕获片段;
2)用上述的试剂盒中的pcr反应液对所述捕获片段进行扩增,得到测序文库;
3)对所述测序文库进行测序,得到待测样本中原发性特应性疾病相关基因或基因突变位点的测序结果。
上述原发性特应性疾病相关基因可选自flg、cdsn、dsg1、dsp、spink5、plcg2、adgre2、tpsab1、foxp3、il2ra、tgfbr1、tgfbr2、was、malt1、card11、wipf1、arpcib、dock8、carmil2、erbin、znf341、ragl、rag2、dclre1c、ada、il7ra、chd7、lig4、zap70、stat3、stat1、stat5b、jak1、il4ra、il6st、il6r、pgm3中的两种或多种。
上述原发性特应性疾病的临床表型可选自过敏性皮炎、荨麻疹、自身免疫性疾病、联合免疫缺陷病、高ige综合征、omenn综合征或骨骼异常中的一种或多种。
本发明涉及可进行快速准确捕获原发性特应性疾病相关基因序列的探针组,有效提高了原发性特应性疾病的诊断效率,可以为治疗提供诊断依据。能够准确快速地检测出原发性特应性疾病的突变位点。
附图说明
图1为患儿编号2021-01-01stat3基因c.1144c>t杂合突变sanger测序图。
图2为患儿编号2021-01-01父亲stat3基因c.1144c>t无突变sanger测序图。
图3为患儿编号2021-01-01母亲stat3基因c.1144c>t无突变sanger测序图。
图4为患儿编号2021-01-02lig4基因c.575dela杂合缺失突变sanger测序图。
图5为患儿编号2021-01-02父亲lig4基因c.575dela杂合缺失突变sanger测序图。
图6为患儿编号2021-01-02母亲lig4基因c.575dela无缺失突变sanger测序图。
图7为患儿编号2021-01-02lig4基因c833g>t杂合突变sanger测序图。
图8为患儿编号2021-01-02母亲lig4基因c833g>t杂合突变sanger测序图。
图9为患儿编号2021-01-02父亲lig4基因c833g>t无突变sanger测序图。
图10为患儿编号2021-01-03rag2基因c.193g>t纯合突变sanger测序图。
图11为患儿编号2021-01-03父亲rag2基因c.193g>t杂合突变sanger测序图。
图12为患儿编号2021-01-03母亲rag2基因c.193g>t杂合突变sanger测序图。
其中,图1-12中箭头所指位置为突变位点。
具体实施方式
下面结合具体实施方式对本发明进行进一步的详细描述,给出的实施例仅为了阐明本发明,而不是为了限制本发明的范围。以下提供的实施例可作为本技术领域普通技术人员进行进一步改进的指南,并不以任何方式构成对本发明的限制。
下述实施例中的实验方法,如无特殊说明,均为常规方法,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
实施例1、原发性特应性疾病相关基因捕获探针组、试剂盒制备
一、原发性特应性疾病相关基因捕获探针的设计和制备
根据原发性特应性疾病相关基因的全外显子序列,对每个区域中非重复区域设计120bp的探针序列,每个序列沿着基因位置挪动设计。利用原位合成技术,大量合成设计的探针。该原发性特应性疾病37个相关基因为flg、cdsn、dsg1、dsp、spink5、plcg2、adgre2、tpsabl、foxp3、il2ra、tgfbr1、tgfbr2、was、malt1、card11、wipf1、arpcib、dock8、carmil2、erbin、znf341、rag1、rag2、dclre1c、ada、il7ra、chd7、lig4、zap70、stat3、stat1、stat5b、jak1、il4ra、il6st、il6r、pgm3。
利用pcr的方法扩增出大量的带有生物素标记的探针,具体方法如下:
将上述合成的探针在总体积1.2ml的ddh2o中,取其中5μl作dna模板,利用通用的pcr引物(5’端序列为gactacatgggacat(seqidno.395)、3’端的序列为ggaacctacgacgta(seqidno.396)),分三管进行pcr扩增,其中引物gactacatgggacat为带有生物素标记的引物。
上述pcr扩增体系:dna模板,5μl;正向引物(25μm),2μl;反向引物(25μm),2μl;mgcl2(50mm),4μl;10xplatinumtaq缓冲液(购自lifetechnologies),5μl;dntps(每种10mm),4μl;platinumtaq(5u/μl,购自lifetechnologies),1μl;h2o,27μl;总体积50μl。
pcr扩增条件:98℃,30s;(98℃,30s,60℃,25s,72℃,45s)35个循环;72℃,5min。
将pcr产物用minelutepcr纯化试剂盒(购自qiagen,货号28006)纯化后,取5500ng,用myone亲和素磁珠(购自invitrogen,货号65002)将pcr产物结合下来。然后加入碱性naoh将没有生物素的互补链变性、洗脱下来;然后将整个磁珠用100℃的甲酰胺液体洗涤,使探针从磁珠上分离下来。用乙醇沉淀后即得到生物素标记的探针,作为捕获探针。
上述生物素标记的探针的核苷酸序列分别为seqidno.1-394所示,且每条探针的5′末端标记生物素基团。
二、原发性特应性疾病相关基因捕获试剂盒的制备及使用方法
a、原发性特应性疾病相关基因捕获试剂盒的制备
原发性特应性疾病相关基因捕获试剂盒,是通过检测原发性特应性疾病相关基因全基因组的突变来进行分子遗传学检测的试剂盒,该试剂盒(规格16人份/盒)包含的成分为:上述“一”获得生物素标记的探针组(160μl)、富集缓冲液bl(208μl)、富集缓冲液hy(800μl)、文库富集结合缓冲液(3.2ml)、洗涤缓冲液wb1(9ml)、洗涤缓冲液wb2(45ml)、洗涤缓冲液wb3(45ml)、文库富集洗脱液(1.2ml)、中和缓冲液ne(500μl)、pcr反应液(800μl)。
1)生物素标记的探针组
生物素标记的探针由seqidno.1-394所示的探针组成,且每个单链dna分子的5’端均标记生物素。
2)富集缓冲液bl
富集缓冲液bl由35%(v/v)人cot-1dna(invitrogen货号:15279011)、15%(v/v)鲑鱼精dna(invitrogen货号:15634017)、0.5nmol/μl引物1、0.5nmol/μl引物2和水组成。
其中引物序列如下:
引物1:aatgatacggcgaccaccgagatctacactctttccctacacgacgatct(seqidno.397)
引物2:caagcagaagacggcatacgagatcggtctcggcattcctgctgaaccgc(seqidno.398)。
3)富集缓冲液hy
富集缓冲液hy由1.25mnacl、0.125m柠檬酸钠、0.1g/100mlbsa、5%(v/v)tween20和水组成。
4)文库富集结合缓冲液
文库富集结合缓冲液由1mnacl、1mmedta和ph7.5、10mm的tris-hcl缓冲液组成;
ph7.5、10mm的tris-hcl缓冲液由溶剂和溶质组成,溶剂为水,溶质为三羟甲基氨基甲烷(tris),tris的浓度为10m,hcl调节ph为7.5。
5)洗涤缓冲液wb1
洗涤缓冲液wb1为0.1%(质量体积比g:ml)的sds溶液a;0.1%的sds溶液a中的溶质为sds,浓度为0.1%,溶剂为柠檬酸三钠缓冲液ssc;
上述柠檬酸三钠缓冲液ssc由175g/lnacl、88g/l柠檬酸三钠和水组成;缓冲液ph为7.4。
6)洗涤缓冲液wb2
洗涤缓冲液wb2为0.1%(质量体积比g:ml)sds溶液b;0.1%的sds溶液b中的溶质为sds,浓度为0.1%,溶剂为0.1×ssc;
0.1×ssc为将上述柠檬酸三钠缓冲液ssc稀释10倍,得到的溶剂;缓冲液ph为7.4。
7)洗涤缓冲液wb3
洗涤缓冲液wb3为0.1%(质量体积比g:ml)sds溶液c;0.1%的sds溶液c中的溶质为sds,溶剂为0.5×ssc;
0.5×ssc为将上述柠檬酸三钠缓冲液ssc稀释2倍,得到的溶剂;缓冲液ph为7.4。
8)文库富集洗脱液
文库富集洗脱液为0.1mnaoh水溶液;
9)中和缓冲液ne
中和缓冲液ne为ph值为7.5且浓度为1mtris-hcl水溶液;
ph值为7.5且浓度为1mtris-hcl水溶液由溶剂和溶质组成,溶剂为水,溶质为三羟甲基氨基甲烷(tris),tris的浓度为1m,hcl调节ph为7.5;
10)pcr反应液
pcr反应液由0.05u/μlphusionhotstartiidnapolymerase(fermentas,货号ep090b011)、pcrmixbuffer组成。其中pcrmixbuffer是由0.2mmdatp、0.2mmdttp、0.2mmdctp、0.2mmdgtp、5%(v/v)dmso、2.5pmol引物3、2.5pmol引物4、4mmmgcl2、500mmkcl、0.8%(v/v)nonidetp40配制而成。上述引物3和引物4最后一位核苷酸均带有硫代修饰,序列如下:
引物3:aatgatacggcgaccaccga*g(seqidno.399)
引物4:caagcagaagacggcatacg*a(seqidno.400)
*表示硫代修饰。
b、原发性特应性疾病相关基因捕获试剂盒检测方法的建立
1、样本文库制备:
提取待测者外周血或组织的基因组dna作为样本;将基因组dna片段化至100-300bp的片段后,采用康为试剂发明的ngsfastdnalibraryprepsetforillumina试剂盒(货号:cw2585m)进行建库,得到dna基因文库;建库主要步骤包括:dna提取和打断、末端补平、加接头、pcr富集和纯化。
2、样本富集杂交
(1)杂交液制备:250ngdna基因组文库、10μl富集缓冲液bl、5μl生物素标记的探针组混匀震荡离心,再加入37μl预热的富集缓冲溶液hy(65℃预热,使用前充分震荡重悬沉淀)。总体系100μl,离心进行pcr,pcr程序为95℃10分钟,65℃杂交22小时以上。
注:缓冲液hy的体积根据所制备文库体积调整而调整,调整比例为hy缓冲液体积=总体积/1.8。
(2)纯化:此步骤所用的缓冲液除了wb2外,均室温放置。
1)取1.5ml离心管,加入50μlmyone磁珠(购自invitrogen,货号:65002),剧烈震荡至少5秒,短暂离心收集管壁的液体,将离心管放置在磁力架上静置1分钟后弃上清;
2)取下离心管,加入50μl1x文库富集结合缓冲液,剧烈震荡至少5秒,短暂离心收集管壁的液体,将将离心管放置在磁力架上1分钟,保持离心管在磁力架上不动,弃上清液;重复该步骤3次;
3)用100μl2x文库富集结合缓冲液重悬沉淀,剧烈震荡至少5秒,将过夜杂交后的所有液体全部加入,震荡混匀,翻转仪上翻转30min;
4)从翻转仪上拿下收集管,震荡离心,在磁力架上静置1min,弃去液体;
5)加入500μlwb1,震荡混匀,翻转仪上翻转15min;
6)从翻转仪上拿下收集管,震荡离心,在磁力架上静置1min,弃去液体;
7)加入500μlwb3,震荡混匀,66℃、850rpm孵育10min,震荡离心,在磁力架上静置1min,弃去液体;
8)重复步骤(7)三次,放置室温2min,震荡离心,放置磁力架静置1min,弃去上清;
9)加入500μlwb2,震荡离心,吸弃上清。
10)加入17.5μl文库富集洗脱液,加完后立即盖盖,震荡混匀,翻转仪上翻转15mm;
11)震荡离心后,放置磁力架上静置1min,将澄清液转至一个干净的0.2ml管中,加入12.5μl中和缓冲液ne,震荡离心,得到30μl模板dna。
3、样本pcr扩增
(1)pcr反应体系
将上述探针捕获后得到的30μl模板dna加入30μlpcr反应液,补水40μl至总体系为100μl。
(2)pcr循环条件
98℃预变性30秒;15个循环:98℃25秒,65℃30秒72℃30秒;72℃5分钟;4℃~1h;
(3)pcr产物纯化:由dna纯化回收试剂盒(cmpure)(购自康为试剂,货号cw2508)纯化,最终产物由65℃预热的文库富集洗脱液17.5μl分两次洗脱,最终得到35μl捕获产物;
注:不同的纯化方法获得的产物浓度会有差别。根据不同的纯化方法,可是适当调整pcr的循环数。
(4)捕获产物经1%琼脂糖凝胶电泳及nanodrop测浓度并做定量pcr。
4、将以上得到的捕获产物通过illuminahiseq2000进行测序,得到测序数据并进行分析,确定是否捕获原发性特应性疾病相关基因或者确定具体的基因突变位点。
上述进行分析确定是否捕获到原发性特应性疾病相关基因或者确定具体的基因突变位点分析如下:
1)snp分析流程
illuminahiseq2000获取原始短序列;去除测序数据中的接头和低质量数据等;把短序列用soapaligner软件定位到人类基因组数据相应位置上,具体请参考:http://soap.genomics.org.cn/soapaligner,html;统计测序结果信息,短序列数量、目标区域覆盖大小、平均测序深度等;soapsnp用于在目标区域找出位点的基因型,具体请参考:http://soap.genomics.org.cn/soapsnp.html;过滤低质量值(质量值<=20)和低覆盖度(深度<=10)的snp;利用ccds、人类基因组数据库、dbsnp信息对snp进行注释,确定突变位点发生的基因、坐标、mrna位点、氨基酸改变、snp功能(错义突变/无义突变/可变剪切位点)、sift预测snp影响蛋白功能预测等;
2)indel分析流程
把去除接头序列和低质量的测序数据用burrows-wheeleraligner(bwa)比对到人类基因组上,具体请参考:http://bio-bwa.sourceforge.net/bwa.shtml;用gatk软件找出序列中所含有的插入/缺失(indel)的信息;利用ccds、人类基因组数据库、dbsnp信息对lndel进行注释,确定突变位点发生的基因、坐标、mrna位点、编码区域序列的改变、对氨基酸的影响、indel功能(氨基酸插入/氨基酸缺失/移码突变)。
实施例2、试剂盒的应用
下面通过具体实施方式对本发明作进一步详细说明
利用本发明实施例1所述试剂盒检测3例样本,结果证实目的致病的目标区域的平均测序深度为300x,远远高于一般的遗传疾病诊断要求(一般为100x)。具体检测实验及结果如下:
1、患儿编号2021-01-01,男,3岁8个月,检测样本为全血,利用本发明实施例1的试剂盒进行检测,检测结果如表1所示,为基因stat3发生突变。在chr17:404
81661位置发生突变,即exon13发生错义突变c.1144c>t,如表1所示,疾病表型为常染色体显性高ige综合征。参考基因组版本为grch37/hg19;纯合/杂合:纯合表示此突变位点为纯合突变,杂合表示此突变位点为杂合突变。
表1患儿编号2021-01-01检测结果
通过一代sanger测序分析,患儿父母并未有变异如图1、2和3所示。该检测结果说明受检人检测到stat3基因有1个杂合突变,已有文献报道,父母均未检测到此突变,此突变为自发突变,提示受检人为常染色体显性高ige综合征,由此可见本发明试剂盒的应用有效提高了原发性特应性疾病的诊断效率,可以为治疗提供诊断依据。
2、患儿编号2021-01-02,女,5个月,检测样本为全血,利用本发明的试剂盒进行检测,检测结果如表2所示,为在chr13-108862784和chr13-108863041-108863042位置发生突变,即exon2发生错义突变c.833g>t和缺失c.575dela,疾病表型为dna连接酶4缺乏症。
表2患儿编号2021-01-02检测结果
通过一代sanger测序家系分析,结果发现患儿的c.833g>t突变来自其母亲,c.575dela缺失突变来自其父亲。如图4、5、6、7、8和9所示。
3、患儿编号2021-01-03,男,2个月,检测样本为全血,利用本发明的试剂盒进行检测,检测结果如表3所示,为在chr11-36615526位置发生突变,即exon2发生纯合突变c.193g>t,疾病表型为重型联合免疫缺陷症。
表3患儿编号2021-01-03检测结果
通过一代sanger测序家系分析,结果发现患儿的c.193g>t突变分别来自其父母,如图10、11和12所示。
以上3例患者,利用本发明实施例1所示的试剂盒,能够准确快速的检测出原发性特应性疾病的突变位点。由此可见,本发明提供的用于检测原发性免疫缺陷病致病基因的探针组及试剂盒的可以为尽早确立诊断和对患儿采取正确、合理的干预以及治疗措施提供准确快捷的检测数据依据。
以上对本发明进行了详述。对于本领域技术人员来说,在不脱离本发明的宗旨和范围,以及无需进行不必要的实验情况下,可在等同参数、浓度和条件下,在较宽范围内实施本发明。虽然本发明给出了特殊的实施例,应该理解为,可以对本发明作进一步的改进。总之,按本发明的原理,本申请欲包括任何变更、用途或对本发明的改进,包括脱离了本申请中已公开范围,而用本领域已知的常规技术进行的改变。按以下附带的权利要求的范围,可以进行一些基本特征的应用。
sequencelisting
<110>深圳市儿童医院
<120>原发性特应性疾病相关基因的捕获探针、试剂盒及其应用
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<400>9
ctcaatcagcgtcttgtctttcaaggggatctcgccattgtagcagatttcccagagcgt60
ggttccaaagctccacttgtcagcagccacactcaggttcttggagtcctcaacacactc120
<210>10
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>10
gacacaccttgcctagacagcaccgtaatggggatgccggggtcactgagcttgatgaat60
gggccacactcactgtcgatgccctcacgggccaggaggaggtttttagtacacacattt120
<210>11
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>11
ggacattctcaccaagtagctcagggcactggccagctgtttggcaactttgaatttcca60
tggtgtggtaaggacatcgcttttccggtgcatgaagagatccagaggacccccttccac120
<210>12
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>12
cctttgaaagagaacacacttactctccacgtcgcggacacagacgccatagaggtacac60
gatgtgtttgtgggagacctgtctcatcatgctggctgcctcgaagaaggcctgtgggcg120
<210>13
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>13
tgccagggaaatatccctgtggctggggtctaagactttgaggatcacttttatcttctt60
ctcttcagaagttccttcgtcatccttgtaatccatcagggtcccagaatagatgtgtgt120
<210>14
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>14
caccctgcagccgtgactcacctgcaccagatccttcttgaggatccgatcgaaactcag60
ctggctcatggggtagacgggctgccactcctgggctttcttagtagccaccagcaggtt120
<210>15
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>15
ctcggggcttgggctggcagcagcgttttagcatgaagctgatgttatccgtgcgcagga60
tctgcttcttgaggtggctcatgaggtctcccaagctggggaagctgcggtccgaaccgt120
<210>16
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>16
gcaggtgacggtcatgaggatgttgtcaaagtcggtgcagctccacctcagcacgtacat60
cccctcctcgcttccttcttgccgcaatttattgatggcgtattctgtactagagggaga120
<210>17
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>17
cagattggaccatgacagccattctgtatgttgtggacgatcaacgggggggccacgtcg60
gtgcagaggtaatgatgggcatctgctgtgagccggaagtagccatctaccagggacaca120
<210>18
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>18
cattttcttgttgtcctgcttgttaatgctgaccacagactcctttattacaatgtgagt60
gatttcagggaagtaagaaaaattgttccactcttcccggatcttgtttttctcctcatc120
<210>19
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>19
tcatgcgtggacacgctgctgtcacaaatggtcttgttgttaaattcctttaggaaatcc60
ttgaaaacattatttatccgcatcctggtgagaaggttcctctgtctgatggacttattc120
<210>20
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>20
ctgatgtccttgggcagttctggcaactgcatcttcttcatcatggcatagtgtgagatg60
gccaggacagccatccctagacactcgttctcaatatcatgtccatcctgctcggtcttg120
<210>21
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>21
ctgagcaaacagatactccagtgagctggcatcaaggagaggggttgcatctggaatctt60
ttttttctcgtagccatttttctgcttctttggagaatgacgccacactgactgctcatt120
<210>22
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>22
gtacttcccatacctcatccggtagtggagccggagggacatcttgtcatcaacggtgat60
ggtgcgatttggagcataccagagcttggtgttctcgtcatacagggcaaagaggttgtg120
<210>23
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>23
ggcatgcctgtgcagccctgatgcacagttcctctgctgtgtactctccactgcccagcc60
ggaggggctccctgtccgacagatagaagatcacttccacccctggctcaggggcctcca120
<210>24
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>24
aagccagatattttccttttttaaaaatatgcaaatctacatacctgcatttattcagct60
gtccagtgttctccaagaagcaaactggattttcttctctactttccaaagctacttcag120
<210>25
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>25
atcattacaggtcatgtagcatttaccacagttgatacacatttcttcatcaatcatagc60
cacaacttgctctacgttgctcaattcaccaaatgttccaaggtactgcagtgcttttcc120
<210>26
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>26
ttcaccaacttatgccaattctcttgttttagatgttaaatcacacttacgttgtctgga60
aagtcagcctttagttcagtgacactttgacaccaatatgcagccgttttctcactgatg120
<210>27
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>27
gaaagttttggtgagggcaaaaccccatccagcttcaaaagctcttcgaatcattgatgt60
gctggtggctggagttgcgctagcaagaccaaaaggatttataaacttcaatccggccat120
<210>28
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>28
tctgctaccgtgactaagatggaagcgtttttggggtcgcggtccggactttgggcgggg60
ggtccggccccaggacagttttaccgcattccgtccactcccgattccttcatggatccg120
<210>29
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>29
gaaccccatggtgaccgggacctcagtcctcggcgttaagttcgagggcggagtggtgat60
tgccgcagacatgctgggatcctacggctccttggctcgtttccgcaacatctctcgcat120
<210>30
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>30
ggattgatgaggagcttctgggagatggacacagctatagtcctagagctattcattcat60
ggctgaccagggccatgtacagccggcgctcgaagatgaaccctttgtggaacaccatgg120
<210>31
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>31
tgttagcttcctcggttatgtggacatgcttggtgtagcctatgaagccccttcgctggc60
cactggttatggtgcatacttggctcaggtaagtagtaagtctagaggtgggggaaaggg120
<210>32
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>32
cccccaatctccctagcctctgctgcgagaagttctggagaagcagccagtgctaagcca60
gaccgaggcccgcgacttagtagaacgctgcatgcgagtgctgtactaccgagatgcccg120
<210>33
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>33
ggcattattgaatgctctgctttcttccagtttcaaatcgccactgtcaccgaaaaaggt60
gttgaaatagagggaccattgtctacagagaccaactgggatattgcccacatgatcagg120
<210>34
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>34
gtacagctatttttaggaattgatccctttatgcttcacaattttattattctgtcttcc60
ttttttagtggctttgaatgaaatacagatgcattatccagaactgaagttgccctactt120
<210>35
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>35
ttactcatagtaatagtatctctgtgactgactaaatcccagttgtttcgatatatcact60
agaatggccacataaacctgggtccttattaatatacgttgcataataccttggatgatc120
<210>36
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>36
tcggtaaattctcttttctggtagactcataatatgcttgagccaacttgaataccatca60
gaagaaactcagtgaagtcaattttcttgttgtggtctatatccaagtgatccatgaaga120
<210>37
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>37
tcttaccttcaggatttgccgaaattccttttccagaagttccttcagctcttttttact60
caatgtgtcagtgtttttatctttttttgaatattgcttgaaaagattaattatggcaaa120
<210>38
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>38
aagcatgctggccgtcggctgcgcgctgctggctgccctgctggccgcgccgggagcggc60
gctggccccaaggcgctgccctgcgcagggtaagggcttcgggcgcacctggagggctgg120
<210>39
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>39
aggtggcgagaggcgtgctgaccagtctgccaggagacagcgtgactctgacctgcccgg60
gggtagagccggaagacaatgccactgttcactgggtgctcaggaagccggctgcaggct120
<210>40
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>40
gctttcctttcagttccccccgaggagccccagctctcctgcttccggaagagccccctc60
agcaatgttgtttgtgagtggggtcctcggagcaccccatccctgacgacaaaggctgtg120
<210>41
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>41
tcagaacagtccggccgaagacttccaggagccgtgccagtattcccaggagtcccagaa60
gttctcctgccagttagcagtcccggagggagacagctctttctacatagtgtccatgtg120
<210>42
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>42
tgcagcctgatccgcctgccaacatcacagtcactgccgtggccagaaacccccgctggc60
tcagtgtcacctggcaagacccccactcctggaactcatctttctacagactacggtttg120
<210>43
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>43
tcaggtcaaggacctccagcatcactgtgtcatccacgacgcctggagcggcctgaggca60
cgtggtgcagcttcgtgcccaggaggagttcgggcaaggcgagtggagcgagtggagccc120
<210>44
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>44
acaggactccagggcccatgaatcactaataatgagcctttcttctgtccgcagaatcca60
ggagtcctccagctgagaacgaggtgtccacccccatgcaggtgagctcctgttcttgta120
<210>45
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>45
tctgcagaaacaggctgatggtgaggaatgtctcttttaggcacttactactaataaaga60
cgatgataatattctcttcagagattctgcaaatgcgacaagcctcccaggtaaggactg120
<210>46
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>46
ctctatcttcaattttttttttaacctagtgcaagattcttcttcagtaccactgcccac60
attcctggttgctggagggagcctggccttcggaacgctcctctgcattgccattgttct120
<210>47
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>47
gttcaagaagacgtggaagctgcgggctctgaaggaaggcaagacaagcatgcatccgcc60
gtactctttggggcagctggtcccggagaggcctcgacccaccccagtgcttgttcctct120
<210>48
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>48
tgagtgatggtgatgttcacagtctctgaagacacatttttactcccaaaaagccccctg60
cagaagtaggagccgctgtctttgagtgtggcttttggaatgtagaagtcagaattatga120
<210>49
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>49
aaaatagtcgtgccagctgaccgggtgttgataaagtagaatagaaatatgtcaaattgt60
ggactgttagttgttaagtggttctggagagtgtgaaaatctgtggcttccagcactgct120
<210>50
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>50
tggtcgataacatcttagtattatcatggaaaagttttgatcttatagacccctcaacac60
ccaaaagtcgtaatcagttctactcaagtaagatttaaatatatatctatattttctggt120
<210>51
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>51
aatctgaatctggaccctgctgaaaactgccgtgatttctcataacactctcctgcccct60
tcagaagtgaaaactggctgatagtttcccaaatgaataattttcgttaatgttaatcgt120
<210>52
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>52
cgttaatgttaatcgtgtgatgttgttgagctagaatgttgactcaactgattcgtgagg60
ggagatcttgcaccagatatggaaccaggacttccaccgaaatcttagctctggaagttg120
<210>53
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>53
gatgttgttgagctagaatgttgactcaactgattcgtgaggggagatcttgcaccagat60
atggaaccaggacttccaccgaaatcttagctctggaagttgatttttttcgtgggagtt120
<210>54
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>54
gatgttgttgagctagaatgttgactcaactgattcgtgaggggagatcttgcaccagat60
atggaaccaggacttccaccgaaatcttagctctggaagttgatttttttcgtgggagtt120
<210>55
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>55
gatgttgttgagctagaatgttgactcaactgattcgtgaggggagatcttgcaccagat60
atggaaccaggacttccaccgaaatcttagctctggaagttgatttttttcgtgggagtt120
<210>56
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>56
tgtcttgctatgctgcccaggctggtctcaaactcctggcctcaagagatgctcccgcct60
tggtctcctagggagctggggttataggcctgagccaccgtgccaggcctgatgcgctct120
<210>57
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>57
ctgatgcgctcttttccgtggttacgttctaccagtgtgacctccatcccttctccctct60
tcacttccttctttctttccttccttgcataaacattgaatgtacggatccctttttggt120
<210>58
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>58
ctttccttccttgcataaacattgaatgtacggatccctttttggtgtttttagacactt60
ctcaggaaacgtacgcattgatttgcaccttgtgtgtccacctgtaaacatcaaattagt120
<210>59
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>59
aggccaaattcagggtatcattcatagcaatcccaaagaaactaactcttaaagaggcca60
attagtaacgcacaggtaaaactttgctaagtatgattctctgcctgggacctcattcat120
<210>60
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>60
tgattctctgcctgggacctcattcatccttctgttcctgacattgcctcatgggtttgg60
ctgccccgttttgaagttacccaaagattattctgccatggcctctgttttttagaaatt120
<210>61
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>61
gttacccaaagattattctgccatggcctctgttttttagaaattcaagactgaaggctc60
agtagaagggggttagcttagaggattgagccaagggtaaaatgatttctgggtagcagc120
<210>62
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>62
tgagccaagggtaaaatgatttctgggtagcagcccagagttcttgaaggatgtttatta60
ggcaacgtgaacgggagaatcagcagtcagaagcggctgagaaaggaaccacgcagacaa120
<210>63
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>63
gggagaatcagcagtcagaagcggctgagaaaggaaccacgcagacaatgtccagttgta60
tagggtagagtgtgtgtgttgtgtatttacgttagtgctggggctttccttgggactcct120
<210>64
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>64
tggagagagttccataccattcatgctttaatgaacacatatacatgaaaataagatgga60
gagttctagtggttttgcccttcctcttcaacggcgaaattgctatttagaagtgggtag120
<210>65
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>65
gccactgccccgtgtcctgtgatgtgacttcagagcttccaaaacgcaggcaagcacaac60
ggatgtctcctgggcgaccatttagcacctttgatttcacttgggcttcatgacttctgt120
<210>66
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>66
cttcatgacttctgttgtctgttcccggcttcttaccaagaaattcttgttcttttggtt60
ttctagattgttcttctactcttcctcctgtagtggtgtaacaaagtcacggtcatatgt120
<210>67
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>67
acttactgtctccgctgccaggtgagcccactcaggaggaggacgctgatcagcaggaaa60
acacagccggccactgtcaatagagagagggagttcagcaaagggccctgggcttggcat120
<210>68
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>68
acaaagccagtgccccactcacctgctacctggtactctgttgtaaatatggacgtctcc60
atggttgcagccatttctgtctgtatttgaaaatctgtgaaaaagagataaagagagaca120
<210>69
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>69
acctgttgttgtgacgaggcaggaagtctcactctcaggacggccttcggggcttgcctg60
aggcttctcttcacctgggggagagagtaagtgatgctggtggagctaaacacaggagtc120
<210>70
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>70
cctgtatccctggacgcactgataataaaccatctgccccaccacgaaatgataaattct60
ctctgtggcttcattttcccatggtggaggttccctgcagtgacctggaagatggaagga120
<210>71
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>71
gctgcattggactttgcatttctgtggttttcctttctttctgttcttcaggttgaggtg60
tcacttgtttcgttgtgttccgagtggctagaaaatatagatggaatgatgcagataatt120
<210>72
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>72
cacagaagggacacttaccagagcttgtgcattgacattggttgtcccaggacgagtggc60
tagagtttcctgtacagagcatatagagtgacccgctttttattctgcggaaacctctct120
<210>73
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>73
ccgggggcacccacaggcccttacctgcctggcagccaggcaccatgatgaacgtgagca60
gtccccacatcagcaggtatgaatccatcttcctgacccttgggaccagccggggcagtg120
<210>74
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>74
gcagtgaagcggaggtctttctctgcagaaggcccagttgccgtcagcctctttttggca60
tcgcgccggaggatgtgggatgggaagatcggtccgcctgggctgtcacccttgtgggtc120
<210>75
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>75
atgggaagatcggtccgcctgggctgtcacccttgtgggtccatccagtctctatcggag60
tcaggagttgctctctttaagtattgggctggcgtgttcagccaggaaactgcctagcac120
<210>76
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>76
tttcaaaccaggttttttttttcaagttcaattgctggaggtgtgggctggggttgatga60
gaaggattgaacttcatttatgaatcctttcttcagctacgcccataaaaggagagggag120
<210>77
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>77
agggttaagttagaacagagaagccagccacctgccaagaaggtctgaagaacccagaag60
cgacattagcaaggggttcgtttcacggaatccaaaggtctgtttcgtcaatcatggcca120
<210>78
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>78
aggcgcctttggcatgcactgcagcgtgtttgtgtttaatctcaggggttctggggcaca60
gcaaaggctctctggggaccctaggctaagaatcttttggaatgtggagcaaaaaaaaaa120
<210>79
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>79
tacttttggaaaattacctttttatctctttaagagatacttggtacattttgatatttt60
catttgcagatatgtagaaatttattcagatttaggagaagagttcactggcatatttag120
<210>80
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>80
gtatgcacctgtgatccagctagtcaggaggctgaggcaggaagatcgcttgatcccagg60
agttaaaggatacagggagtcatgatggcaccaccgcattctagcctgagtgacagagtg120
<210>81
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>81
tgtagttttgttttgtctttattgatagaatgtaatgggcctttgggatcttaagtacta60
cttatgttacctttgttattcagatgctgaacttaagcagtgaaataatagaaactaaag120
<210>82
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>82
agtctcggtgaactgttctagctctcttcagttttcccagtggtttatactttggctccg60
tactttgggaagaccggcataaaggctttaagctgaaatgaatcagaatatttgattttt120
<210>83
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>83
aaatacaaaaaggcagagaaataagatttgaagacgtcctacccctttcgaaggcctcac60
tgccaactgcctcctcctggcagattccgggaagcctttaggatgatctacaacatacca120
<210>84
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>84
gtcccacaaccagtgactactcacatttcgacaactttatccatagttgtgccaactgga60
atgacatttggatatgcgttcacaggacagaggtagctcaagaaatctttaatctagaaa120
<210>85
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>85
catccattactaagttgataatgagaacacaaagacttttatgcccttctgcaaatggac60
agactagaacccaatgtaattattagtatcccaccaaactgaaagaggcagaaacccaag120
<210>86
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>86
cagaacactgagcgataagggttatgagtatatgggatcctcttacctcactgtaggagg60
agtgaaaagaaaaacaagctctgtatgaactctctccagtcctaaagggaagtgaaaaca120
<210>87
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>87
tttctgctcctttctccaaaccacatggtggatggcttaatgctgattatgtggagtgga60
attctatttctggaagtaattccacagggtaatttgctccactgaaaatattcctctgcc120
<210>88
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>88
ggaatcacttgcacacgtaccttgggatgccggcatgcatggatctgagtgttgcggtct60
gttgtgagatgatgaaggatctcaggcatgttcctaaacatgtctagcttattcacatga120
<210>89
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>89
aacagtcaccatacctggactcctaattcttcactaaggttggtgaacagatattcatag60
ccataagccgctttgcagttcagccacacaacatggtacgggctccgagtgatccagctt120
<210>90
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>90
cccagatgataaccctgttcctccaggcagacttacccgacttggaatttggtaaaatct60
tggatcacagaacgtagtatccaaatatacactttggatgtctttgactctgaaaagaaa120
<210>91
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>91
cggagtgcagaagctccattctagcagcttctccttgcgccaatctgaagtctcctgtgt60
acaggacagttccattattgccctgaaataaaaacctgaaaaagaaatatatcccagtga120
<210>92
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>92
aatatgtgtctacatataataaaatgacaaaataaatgacccccttacataactgatccc60
ggacagtgaccagctggtaagagagtcacaacaatctcttccttctaaaaagaaaataaa120
<210>93
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>93
atttaatatttagttacctctcctgatgcttcatccactaaagatatctgggtaggagtc60
tcgatttcaatagatatctataaaaataaaataagagaccatgtatatagcagtttttca120
<210>94
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>94
agttattaaaaaaataaaacttacaattcgtttcttccaaaatctgtatttcgggctcgt60
taacaacaactccttagtcacaggtgaacagtatagataaaccttcaagctgaaaggaaa120
<210>95
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>95
aaataaattacctgcactccaaccttcttttcaaggtaggggctcttaatcctttcatgt60
gatctaaaaacaaaagaacaaaaactcatgaatatgtttttccagaggattccttagccc120
<210>96
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>96
cctcactcacctttgtggcagtgggacaggaagtaggcgcgggccctcaggttctcccta60
tcgaagcggtctatggagatagttggatactcggccatctgcccctcgaaagaactcata120
<210>97
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>97
ttagaaagaaaacagagttcatagagaaagttacaaaatagattttcttaatacctgaaa60
cttggtaaaaagctgtggtattaaactgtggtgatgtcattaggggagccctctacccaa120
<210>98
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>98
ccctctacccaaacagaaaggaaggctcaacagagcacaggtgtttgttgtaaatggaaa60
aataaatccgtcttattttcatactgcagcaagttaacactatcgaaagcattacagatt120
<210>99
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>99
taaactagataattaatgtaggatgcaaaagatagggcctcatagtatatattaaaaata60
ttatataattagcatgactactactgctgctgctaactactactactactattattgctt120
<210>100
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>100
ctactactattattgctttgttggtgctgttagtgatcatttttagtgattgcccacaaa60
aggaacaatactggatatgaggaaattgatataatttatcttttcaagcatactgtgggc120
<210>101
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>101
tctctgcagcattcctgggggtggccacagaggcctggaacttgacacacaaggccttca60
ctttttcaaaggtcattatttcaccattggtcaggaagaacttgttcccaacttgtttgc120
<210>102
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>102
caaacttcaagctgcctggagagtaagagaaaaagcttacactttttgatacgtgccatt60
tctgtttgcagagcttttctttctgaggcagccaggctactatcaccatctagaaaatta120
<210>103
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>103
tttccagggtctcctttttggccctttggtcctggtgacccagaaggccctggatttcct60
ggaggccccaactttccaggggggccctgtaagcctctgagcccttggcctgttggaaga120
<210>104
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>104
gagagctacaggcaatcactgcagggcaggtcttttgggcatcctcacaggtcacagttt60
ctgagtaagacgctgccaccatactcaggagaaggagagggagtgatggaaacagggaca120
<210>105
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>105
taagacgctgccaccatactcaggagaaggagagggagtgatggaaacagggacatggtc60
ctcaccttggtgtgagaaaactcagggaaggttaatctcagttaatgaacacatatttac120
<210>106
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>106
tggggtgtgtttggtcccatcttttcctctgccacaacaccaccatgatgatcaccataa60
catcatatttattggccctttatcaagctatgtacatggtgagacaggtcaggacaggcc120
<210>107
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>107
ttcctctgccacaacaccaccatgatgatcaccataacatcatatttattggccctttat60
caagctatgtacatggtgagacaggtcaggacaggcctccatttgtctcaggggcaaagc120
<210>108
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>108
tgatggggctccagttaaggcaatgaaggctttgccacaccatagagggctcaagggaag60
ggtcctaaaagaccagccctgagcagcctggtgggaacagcctcttggccttctgcccag120
<210>109
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>109
aggctttgccacaccatagagggctcaagggaagggtcctaaaagaccagccctgagcag60
cctggtgggaacagcctcttggccttctgcccagctcctggctgaagctgtgatctgtgt120
<210>110
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>110
gcctcttggccttctgcccagctcctggctgaagctgtgatctgtgtagctgtgactgca60
gggcttgagaatggcggagggcttaggcagtttggacagggtgaatcgggattagcgtgt120
<210>111
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>111
ctgaagctgtgatctgtgtagctgtgactgcagggcttgagaatggcggagggcttaggc60
agtttggacagggtgaatcgggattagcgtgtaaacccagccacctccctgaaaaacagt120
<210>112
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>112
tgcctccgctctcacccctcagggacctcagtgctctgtataggtattttctcttttcct60
tttcaaaataacatttttttctttccttttaattttcttattataaaagcaatgatcctg120
<210>113
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>113
ggaaaggcagggttcagtactcactagacctaaatggcaccctgctgggactctcaggca60
tggtctcactcactttctctctacttgatgtctcactgggcactcccagatttgggacct120
<210>114
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>114
tataggcctggttcagcccccacattgtacagatgggcccagagaggggaaaagcccagc60
caaggggagcccctttttagaatttccctaaggcctaatggggatctaggtggatggctc120
<210>115
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>115
ctctctccttacctggaatcccgtatagagaagcggctacacagatggatatcctctctt60
caccgaggctcctggaatggtggcatcagccccccaggcagcccactgtggctccttgca120
<210>116
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>116
ggaggctgggaaggacagtggaagctgataaacagctcagcagcatgttctgagaaacaa60
gagggcaaggagagagcagagaacacactttgccttctctttggtattgagtaatatcaa120
<210>117
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>117
ccagaagtttatgaatgctcataatgcattaaaaacctctgggtttaccatgaaccctca60
ggcaagcttaggggacccattaggcatagaggactctctggaaagccaagattcaatgga120
<210>118
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>118
gccaagattcaatggaattttaagtagggcaaccacttatgagttggtttttgcaattga60
gtttccctctgggttgcattgagggcttctcctagcaccctttactgctgtgtatggggc120
<210>119
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>119
agaaactgatgagctgattgcttgaggcttttagtgagttccgaaaagcaacaggaaaaa60
tcagttatctgaaagctcagtaactcagaacaggagtaactgcaggggaccagagatgag120
<210>120
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>120
gagctgattgcttgaggcttttagtgagttccgaaaagcaacaggaaaaatcagttatct60
gaaagctcagtaactcagaacaggagtaactgcaggggaccagagatgagcaaagatctg120
<210>121
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>121
agctcagtaactcagaacaggagtaactgcaggggaccagagatgagcaaagatctgtgt60
gtgttggggagctgtcatgtaaatcaaagccaaggttgtcaaagaacagccagtgaggcc120
<210>122
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>122
tgttggggagctgtcatgtaaatcaaagccaaggttgtcaaagaacagccagtgaggcca60
ggaaagaaattggtcttgtggttttcatttttttcccccttgattgattatattttgtat120
<210>123
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>123
atttttttcccccttgattgattatattttgtattgagatatgataagtgccttctattt60
catttttgaataattcttcatttttataattttacatatcttggcttgctatataagatt120
<210>124
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>124
caactttgagaaaatcagtccttttttatgtttaaattatgtatctattgtaaccttcag60
agtttaggaggtcatctgctgtcatggatttttcaataatgaatttagaatacacctgtt120
<210>125
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>125
ttcagagtttaggaggtcatctgctgtcatggatttttcaataatgaatttagaatacac60
ctgttagctacagttagttattaaatcttctgataatatatgtttacttagctatcagaa120
<210>126
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>126
ttatttttactttttcatttcaagaaatttagagtttccaaatttagagcttctgcatac60
agtcttaaagccacagaggcttgtaaaaatataggttagcttgatgtctaaaaatatatt120
<210>127
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>127
catgtcttactgaaacattttgccagactttctccaaatgaaacctgaatcaatttttct60
aaatctaggtttcatagagtcctctcctctgcaatgtgttattctttctataatgatcag120
<210>128
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>128
ttatggggcaggggtaagcaagtttttaaatcatttgtgtgctctggctcttttgataga60
agaaagcaacacaaaagctccaaagggccccctaaccctcttgtggctccagttatttgg120
<210>129
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>129
tttgtgtgctctggctcttttgatagaagaaagcaacacaaaagctccaaagggccccct60
aaccctcttgtggctccagttatttggaaactatgatctgcatccttaggaatctgggat120
<210>130
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>130
catccttaggaatctgggatttgccagttgctggcaatgtagagcaggcatggaatttta60
tatgctagtgagtcataatgatatgttagtgttaattagttttttcttcctttgatttta120
<210>131
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>131
tgatatgttagtgttaattagttttttcttcctttgattttattggccataattgctact60
cttcatacacagtatatcaaagagcttgataatttagttgtcaaaagtgcatcggcgaca120
<210>132
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>132
aaagagcttgataatttagttgtcaaaagtgcatcggcgacattatctttaattgtatgt60
atttggtgcttcttcagggattgaactcagtatctttcattaaaaaacacagcagttttc120
<210>133
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>133
tgtttttttggaaatgagtgggccactaagccacactttcccttcatcctgcttaatcct60
tccagcatgtctctgcactaataaacagctaaattcacataatcatcctatttactgaag120
<210>134
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>134
gaactattttgaatgcatgtgactaagagcatgatttatagcacaacctttccaataatc60
ccttaatcagatcacattttgataaaccctgggaacatctggctgcaggaatttcaatat120
<210>135
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>135
ccttaatcagatcacattttgataaaccctgggaacatctggctgcaggaatttcaatat60
gtagaaacgctgcctatggttttttgcccttactgttgagactgcaatatcctagaccct120
<210>136
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>136
tatactccagggaaattcaccacactgaatcgagcatttgtgtgtgtatgtgtgaagtat60
atactgggacttcagaagtgcaatgtatttttctcctgtgaaacctgaatctacaagttt120
<210>137
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>137
gcattacatcaaacagagttcaaattccacacagataagaggcaggatatataagcgcca60
gtggtagttgggaggaataaaccattatttggatgcaggtggtttttgattgcaaatatg120
<210>138
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>138
gtaatctacataaatatacctcagaaatcatttttggtgattattttttgttttgtagaa60
ttgcacttcagtttattttcttacaaataaccttacattttgtttaatggcttccaagag120
<210>139
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>139
tacaaataaccttacattttgtttaatggcttccaagagccttttttttttttgtatttc60
agagaaaattcaggtaccaggatgcaatggatttatttgattcaggggacctgtgtttcc120
<210>140
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>140
tgtatttcagagaaaattcaggtaccaggatgcaatggatttatttgattcaggggacct60
gtgtttccatgtcaaatgttttcaaataaaatgaaatatgagtttcaatactttttatat120
<210>141
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>141
gatgcaatggatttatttgattcaggggacctgtgtttccatgtcaaatgttttcaaata60
aaatgaaatatgagtttcaatactttttatattttaatatttccattcattaatattatg120
<210>142
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>142
ttcaaatgactgactttggcacatcattcatgcatcatataatggggaggggaaatcctt60
aaagaaaatgcatacaagtaattcaggaaaacatgtcaaacttgatgtagattatttatt120
<210>143
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>143
taacttgttcatgggcataatccaaagggttgcttgaaggtatataaagaacactaatgt60
gggatgtagtagatcttgcttttaagttttggccaactgccgaattgttatgttgcacaa120
<210>144
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>144
aaaataatcaatttaattcttttgggtgttaatttctttattggctaaatttgtcataaa60
cactttataggttatttgttaccaagacttatataataaatataaacatacctcgatgat120
<210>145
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>145
agaatgactttatttatcattgcattataaacacttttttctggcccttaattcatgtaa60
ctaaaagaaaacataggcattttaaataaacaaaaatgtatttttaaaatcaatgttatg120
<210>146
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>146
taaacacttttttctggcccttaattcatgtaactaaaagaaaacataggcattttaaat60
aaacaaaaatgtatttttaaaatcaatgttatgattttaaaaatagattcaaaaattcca120
<210>147
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>147
ggcattttaaataaacaaaaatgtatttttaaaatcaatgttatgattttaaaaatagat60
tcaaaaattccatacacctgaatctgaaaggcttttgcaaaactaatcaaacaaccttct120
<210>148
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>148
ctaatcaaacaaccttctaagaaaggatttcttggcaggagtcaagatttttccagaacc60
ttttttacggagggatttcattggaggcttttttaagggtaggactctttggggagtgtg120
<210>149
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>149
ttcccttaaagtttactaacataggctcccattttttagctttggtagaaatgaaggtat60
cagacctacctgaaggccagaggggctgctcacgcctctctgaatctttgccgctaaacc120
<210>150
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>150
ttcactgactgcccactatgactgtggcaagcagaaggctgactgtaaagagagtgtgta60
tgtgtgcctacagatgttcttatgaacactgatctaatgggagtaactgactgtgacctt120
<210>151
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>151
cctggtggacatggtgaatgacggcgtggaggacctccgctgcaaatacatctccctcat60
ctacaccaactatgtgagcatctgcaccagggttgggcactgggggctgaacaaagaaag120
<210>152
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>152
gaggacctccgctgcaaatacatctccctcatctacaccaactatgtgagcatctgcacc60
agggttgggcactgggggctgaacaaagaaaggggcttcttgtgccctcaccccccttac120
<210>153
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>153
attagaattatatttatattgaagtgcacagaatgttttttcctcttttgtcctggtatt60
acatttagaaaatttactattaaaaaattttaaaaggcgataccttgatacttcattgtt120
<210>154
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>154
ctccattttctgcacattaattaccccaggaagaggggaggagggatgaggcatgaaata60
aaaatgctcgggaatataggtgggtatgtttcctgtatcctcccttactcatctctgtct120
<210>155
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>155
atcctgaagaggcttgtagtgagagatgtttctcagcaacccagggccacctacaaggcc60
tggtaagtaatctgagaggaaataagcttcttaggatcaggattcccccatgttttcaac120
<210>156
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>156
ggagccccattcaccttttcagtatatagagttatggtctttggagagacatgagacaca60
gagtaagagggacggagtgggctgtgatgactgctgcccctcctcaagactggggcacag120
<210>157
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>157
gagttaattgaacattttatgacaagatttttaggaaaagatagaggaatagtagtacgt60
tgtttcaaaataagaacaacgatagcaaaagagctgagaatctgctatagtactatgtct120
<210>158
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>158
atgttcctgacttcagataaaatttcattactatatttagttatggaatgataggaaagg60
cataaatttttaaacttaataaaaactggaaatatgttaagaattaggagagccttacta120
<210>159
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>159
tttctattgtattctagttctctaacctttgcacacatttgtttaggggctatggacaga60
gagattctcctgtctttgttgtcagcattgtgcttagagaaagtagaacaatctgagtca120
<210>160
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>160
caatctttaatgaactggtgagaattagaaaaggaaagtgagagtgatcaatgctcagag60
ccagttccacagcctgctgaccttgggggaagaagggacatccgtaaatgattatggagc120
<210>161
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>161
aagcagacgttccgttcttggcattacgcacctgctaaagggagagagagtgtttgctgt60
ggtagaccagacaggggaatggaaggcctgataattagcccatcctctgacaaaccctgg120
<210>162
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>162
gttgcacctcttgcatatagagagagactagatgctaattaaatataaagtacttgtgac60
tatcctcccacataacactcttttgggacaccacaaatttcttccccatctacccactcc120
<210>163
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>163
cccactcccacctccttcccccagcccttcaagcagaacagtctgtgtatactttggaaa60
cataaactcacattatcattaatctaagccaagcatcattgagaaagtaaatctgctgca120
<210>164
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>164
taccaagctcaaagaggttgctatcagataggatttgaatgaaaaatcatgtgatggtcc60
tgggtttagttaggcttttactgactccattcatcccgcaacaccccgtgacccctctct120
<210>165
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>165
gagttgtcatggcaattatgattgacaatggagcatcctgtgaatgccaacaatgatccc60
taaaatgacctctgccctacaggctgggtcctgcagggaatgttggcaggctgggctcaa120
<210>166
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>166
ctctctctctctctctctgtccacacacacacacacaccggtgtagttgctagggcagag60
atagctccatgaaacaaagtataaagccaggtctgcctgggaggtccaaccttcccaatt120
<210>167
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>167
attaatttataaatcagggcaactgcaacaggcaattttcttatttggtctaccacttta60
gagcaatgaacttgacttcttttttggctaaccaaagaacaatccaaagcagatgtaaac120
<210>168
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>168
ccaggctacgagctgggcagcaattcttcctggcaagtagttcttgcaatttgcctatac60
ccagagcagatgctgcttctgcttttcccaaagaaagctttctctatattctgcctttcc120
<210>169
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>169
attagagagggtaggaaggcaatgaaaccactctgaattatgtaaagatcttgctacttc60
ttatagcctaataggaagaaatccaggcaggggttatagggcatcagctgttgaatgaat120
<210>170
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>170
agcatggtaaacaggtgggaggagaaaatgatcattttggaattgtaacgaagtctctga60
aatggaggcccttggagtgccagcttcagaactgtttggccagaaccaaggacttaggat120
<210>171
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>171
gccagctcatcattgctgttgtttatttattcatgctcagatggggcggtccaagcccgt60
ttgttcacagacatctttcccctcgttgaatcttgctctctgcctgcctttcctactagt120
<210>172
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>172
ttccaccccaagggagcttttctccccttgaggccaagacagggtggggcgggggacagg60
gatgggcagcacataaccctctggccctctgcccttcccatgcaggtggtcagcactggg120
<210>173
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>173
ccccaaacctctgatgcttccaacgtggccctccctgtccctgatgcctgcacctggcgg60
tcccctactgctgcctgtctaccctgtcccctaactgcactggtggagagagcagagaat120
<210>174
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>174
cagtggcatgtggtgggcatcgttagttggggctatggctgcgggggcccgagcacccca60
ggagtatacaccaaggtctcagcctatctcaactggatctacaatgtctggaaggtaagg120
<210>175
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>175
ctgattccagaggccatcaggcctaaaaggagtagtgcaggaagctgtgctcccatggcc60
agtcccagattcaggtacatacgtactgagctattttctgcagatctctggcctaaggcc120
<210>176
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>176
ggagacaggagaaaaatggaaaaagtcatcctcaagaagattaaagtctaactggaagca60
atgactatctgcttgtgaagcctttttttttccaagttagacaagtgttaactacagcta120
<210>177
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>177
aaagctgctgaggccagcttggggccccttctttcctgtccctgggcacttccctggctg60
ctctctttccccactgcccagcccaaggagtcccctctgcagctgacccgggttcagcct120
<210>178
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>178
ggggcatgaaggatcttctgttttttcgtattttctgttttttttccttttagtattcag60
taaaggccttttttcagagttcccacctgtctcaggtaggccagagagagcaacatggtg120
<210>179
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>179
aggtattcagtttcccagaccttttgctcctagaaacaggtcctagagcggagggcaagg60
gtgtggaagagactgggtgggtgagaataggctgggccagactctgagcctcagccctgg120
<210>180
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>180
ggctgcaggtctgtttatagcctgtgtgtgctgggaaaagacatcagaagtccttgccat60
tcccccatcccacccaagccctcagagcctgctcccatttgggtctagagcagcatgttc120
<210>181
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>181
cctcactgggttggctgaagatagagaggaggagccagggcctctgcttggggtgagatg60
ggagatgggggaacaaagggcagaggagatactttctcctccggacacttttgctgaacg120
<210>182
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>182
aagtggcctggctttgaaacatggtgctatcccctagcaagcagaggaggagcttccaga60
cagaaaatggtacctttccccagagtctatgttgaggtgcaccagcctttccagtcctga120
<210>183
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>183
gggcaaagagcatgacctgaaaactggctgtgcctcaacgtcctcaactgtaaaagcagc60
tgacaatggctggagtggagaggggctgcagctgtctgggtgtggtgttccctctgtttt120
<210>184
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>184
actgaactccaaccctgcacaggccccagcattgctcctctgagagccaggctcagctca60
cttctcaggcataccaagatggccacgaaccctacctaggtgctctcaagccccaagagg120
<210>185
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>185
gattataacgttctgttacataaaatagggtatgccttacatttagtaaatccagtatga60
acaaaaaacttatgccacagttgcaggctgggtaaattatgggaaggaaggaaggaatca120
<210>186
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>186
tctagttgagagatgaggaaactgaggcatggggatttcagggctcagtcgaggtggcaa60
gggaagttctgctcagcatgggcatattagacacatcagcaggttattgcagcaattgaa120
<210>187
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>187
ttcagttggggcagctaacttgctgtcaatttgtctaagccggaaagggagaatctgcag60
cattagtctgtggcatgtgagaggcccattaataaaagagccacatgtgctcaggggaaa120
<210>188
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>188
ggacttcccttccaggtgggactgagctcaagggtgatgttaattactgtgtatacctac60
cctagtctgatgtgtcctcccaaagactcatcaccccaaaccggagagcctctcatcttt120
<210>189
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>189
tctgtgcaggaagctgcagcaggtaggatttgtctccctgggtccaggatgtgactgtca60
cccagagggcacctgaactgccccaactctgcttcctcctgtagccagtggtctttttcc120
<210>190
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>190
tgctttgcagctccacctgcatttgctttgggacccctttgagtccctcagcgcttctct60
ttcctacatcctcccaggcgatcacccccttcggggcttcccttcctgtttctctgtctg120
<210>191
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>191
gattataactcctgaagcagcccgggtggcactttcggtaggtccagggagttaccaaag60
taatgtgcctgtattccttgggggccagtttcattttatttgacaacttaagtttccatt120
<210>192
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>192
ttttcagatcaactcaaggttctcgaacacatctggtacgctcttaactttagctgctag60
aagtattcccagcacttggaattccatcccatctacatcagagctgtcaacaaaatggta120
<210>193
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>193
caagcatagaattgacccttaataggactagtcatgatggacagagatccccaatgataa60
aggtattcacaaagaacacaggttacaacccaattcttgagggacccttgagtgagccag120
<210>194
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>194
gtggtcagaagagaataaaagcagcaaaaataatgtgatcagtagcagatccctttcctc60
ataagctgtttctttcagaataaaaaattcactaccaccccaccccaaaccacccaacac120
<210>195
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>195
gtttcccagtcattcctgacaagagtttcacgaatgaaatgagtatttgtataaatgaat60
gacaagcaggtatgggccagaatcacttcatgactattgctgtgtctagtaattgttatt120
<210>196
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>196
gggtttagggtccactccgcatactcccacttgcccttggcattggccccatgggtggac60
catggaggtgggtgctgtatggcaacagtcattcctcagcagagtggagagggagacaca120
<210>197
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>197
ggattttgtttatccgttgtgcttgttctccctacatcagtgatgataaaatattgtcaa60
aataatatttttccatactataaaggcaaaataaaagcaaggacatatgttctaataaaa120
<210>198
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>198
ggaaatggccccaggaccatcaaggctttgccctgctttctgggcagggttcggcaaaat60
taaaaaggaagaaagattatttatcgtccacccagtgcaaattaagaaaaatggtatggt120
<210>199
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>199
attagagtgtcggtcccaaatctcgcctacctggcagagattcttatcgcggttagtgct60
ggccaactccctcccctgctagatgggattctaatccctataagccctggaaatgaacag120
<210>200
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>200
actcatcttttttccctgtagaaatgaaacagatctattgtctgtgtttccttcttctgg60
gcatcttaaatgatggataggttcaactggctgagagttatccttgtaaataagggtatt120
<210>201
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>201
aatactgctagttttttttttaaatcaataaaagcaacactgatacattatagcattcca60
aggaacaaaacagagggagaaatttcaggttcacatccaggtcagtaagtcagcaattcc120
<210>202
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>202
catagcatacaatcttttttatttttctagtataccatcttataaattttctttgattcc60
gccatttattgcctgagtttcaagaaataagcaaataactgctatagagccttcaagtaa120
<210>203
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>203
ccctcctactctatgtaccctgccatgctgtagatgtagaaaggcaggttcttattgctg60
tgtcatcagggagatgcatcctattttcaggtgagacttgcctggagttttggtgctatt120
<210>204
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>204
cccttggcctgtgctgacacatccttagcccagagggtgagcttcaccaacgttggcctc60
atatctcttgtctgctttctgctaattcttttatcctacactagaatcacaatatctatc120
<210>205
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>205
tgacttacccatggtagaatcttactaaaccagatatagcaccaacttagcctatggagt60
gaaggctggttctcccatatgcaaacgtaactcttaagaagtgatagaataagggagaaa120
<210>206
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>206
tgcttcgatggcagaaatattcttggagaagttataaaatcttaacacaaaagcagaagt60
ccatcttcaggcaagtccaagtgaaaccaccgcacacaagcctgtgtgtttatgaggatg120
<210>207
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>207
ttcgtctaccccacaggctgccgctttgacctgataggtagaagtcattagcctgtgagc60
ttatttaagcaattgggatctgctatgtcagaccacccaatgaaagaaatgtctcctctt120
<210>208
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>208
tggggtagggagggtgaaggagcagagaatgaggacaaggggctgcccgcggggccctaa60
gctcctcccctgaggtgcacacgttcttcctctctcagcctctttggatgggctgcctcc120
<210>209
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>209
gtgctggattgtagcatggtgcctggtacaggatgggcacagggatgcatgggagagcag60
cctcgcttcagccgaagggaatcaagatgttagaaatgggcacagctcctcctactgagg120
<210>210
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>210
ggatcctggggatggcccacctgccttggctttgaaggcctggcccacactgggccatgg60
gatctgctatgggcaggctggctgggagtctgctatggcttcacacattgtctacttact120
<210>211
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>211
accctttgaaaggtaaaagctttttgttttataaaatctgatttacgggcccccggtgca60
gggccaaggttttatatattatgcaagatttcaaaccgtggctgtaaaacgtacggtttc120
<210>212
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>212
tggctcaccgaggcgcgcagggagtggaagtgcctgaggataaaggtgtacacgtgcgag60
tctgagatgatggaattgctgctcaaacttcgtccccttctctgccctcagcaaacttat120
<210>213
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>213
tgcatctcgacacccactacggccaacacacagcccgggcccctaggtgatgggtggaca60
aatgagatgcggtcaggattataagccacacctcgcctgactggttctccacttcctttt120
<210>214
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>214
aggactgaggcctcctgggcctcagaggcccaggcccagctgagcacgaatccctggtcc60
accatggcttagcccctgctggcaaaagggcaagatcaactgcccctctcagaacccggc120
<210>215
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>215
aagggatgtatgggagaagccagggagtggagagagaactcgtgttccccgatacaaggc60
tttttacccatgtctccccaaatagccccccctcctcccactagccccgaggctggagat120
<210>216
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>216
aagggcttgagacctacaggataggcagggggcccagcagttttagggtcccctcaaagt60
ataggtcttttgaatccttgatctggtgacatttttcattatatcaaccttcatttctgt120
<210>217
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>217
cacgtcccttgcctttgcagaagcctctcaccagcacccttcctgccccggtgacttgct60
gccagagctctgcctggaagtgtgatggaagccacctggcatgagcttgaaaggaactgc120
<210>218
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>218
tatttcccagatttgaacctggatcttgttgaattggagaagtgggacagacatggtaca60
ttttgaaacccaaccattatatccttaaagagaagtacatttctcccttccactttgttc120
<210>219
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>219
tactcatttataccttactttccaatagttaatgttttccaaatacactattgtcattct60
tctagcaaaagcccagaaagtattttatagggagtcccatggaagataagatttgatatc120
<210>220
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>220
atagagttggcatacctaaattctagaaatgctaaattgcaaaatgaattggaaacaatg60
gcatttctttggagacaaaaggtattgaaatgggacacagccccatttggtctcttgaca120
<210>221
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>221
attttcttaaagtcatttaagtttattatttcacagaaatgaaataaactgtacagcatt60
ggtcataagcataggtactagatgagtgatataaacaatgataaatggcaggtaattatg120
<210>222
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>222
tctgtactggggctgtgaaaggctgggctgccctcacactgcaggtgtcgggggctcagg60
agcctcttcttatcttttgtgtgactgcaactattttcccaagaaaattaaagtgactta120
<210>223
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>223
agtttatatccatatatctgtaatggtgggcacagtgtagggtgagaagaggaagagtag60
gagagattatttaaaaagaaagttttacaaccaggaagtattggaagaggcaaatcatgg120
<210>224
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>224
ggcacagtgtagggtgagaagaggaagagtaggagagattatttaaaaagaaagttttac60
aaccaggaagtattggaagaggcaaatcatggcaactctggatgtttttcctaaaacccc120
<210>225
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>225
aactgggggaaaagccactcattctcacaacaatccaatgccccagagcccatgggatga60
aaactgtgacttaaaattcagcatagaacacagaataaaaaagatcagaagggcatgaga120
<210>226
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>226
gcaggacgaggcgctgagtaagggacggggctgctcccagcagaatccacaaagtagctc60
attctgctctcagcacctgtgtgagagaaggcgttggctgagcctctggatgctcacagg120
<210>227
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>227
tgcgtgacctccacaggaaagtgcaactatcttccaaaaatgtataaattatcaaagatg60
tattctataacaaaaatgacagtccttgagatgtggtggattaatattgctagatcagga120
<210>228
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>228
acaccacacaagattgtaggagggatcaacctcagctcaagagtcaaagagatgagaagc60
atcagagcagcagcgtagcttggagaccatacaccaggttcctgacaccagtccccgggg120
<210>229
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>229
tatgggcacttacatagcaagtcagcccctgtctcatgcagagcttcaatttctagcccc60
ttatcatcagcgatcatactattccatggtttctaattttgacactttctatttttgtac120
<210>230
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>230
ctgggcttctggctcctgtgttgaggggaagagagtggggagaggtcggaccagcattca60
actcacaggctcttcccagcttccatcccctttgccctggagctgcagaatttcctgacc120
<210>231
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>231
atatctgaaattacctcgctaaaggacgttatcttagtagtagaaattaatattttcctc60
atgtacggttacccacttctcattgcctcatatctgtaataaagtcagactgcaagatat120
<210>232
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>232
agggagttgccagcacttgccgggcaggacactgtagagcagctccagccagggctggcc60
tggtcggggagcaccggtgaggagggcctgctcctgaaggaatctctggaccctgtccca120
<210>233
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>233
aatcctacatgaaaatccttgagagttcaaaaggaaccaacattcagtgcgagttctaaa60
gaatgataaaccttgatgtttgcctttttctttttataaagagtagcagctggttggctg120
<210>234
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>234
agctaggattgtcctgtaggtggggcttcagctgctggagtggaccggaaatctgccttc60
tccaggggctttctggagccttccctctgccttggtcctgagaacacctggaaattcctc120
<210>235
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>235
ctgttgcccttggttcattatccccttaccattcagattctgggctagccaaacctcatc60
ttcccagctcatcctaacgaacgccctcggctctctgcaggtgactgcctcctggaccaa120
<210>236
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>236
gccctgtctcccagaagcttaaacatctccactgtaatgtaacaaggcctttttcttcac60
ctcactgaacaaagggagctacagaggggagaaagggatgtcaaagactgcgtgtgagct120
<210>237
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>237
agaagacttttgtaagatcacttgaaattccctaggaattgcttacagcgctatcataaa60
aggaacaaaaaaatccatgtgaaaatattaatcataggaggctggaaaggcaaagacatt120
<210>238
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>238
tattttggcattttgcaggctataaaatgaaaatatagctgagaagcacaaagaattctg60
gcccaggtcccagatgagagggaagggtggcttgagaggtgaatgtatttgttgtgtgaa120
<210>239
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>239
gtcatggggcgcaggctcacagggcggctcagctgctggggacaccgccgtcagggatgg60
ccctggcacgtggcacgtcatgccgcaggggatctctgcatcccagcaccccacattcct120
<210>240
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>240
ctcctagcttttaaccagactgttaatgcctagtacctagcaggcaatcaataaacacat60
gctgattgcaactagagaataatttaattcaattattcagaaatttattgaatttctctc120
<210>241
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>241
tgagtgctagaagtggaaatgggggtggttgtagctttgctgattcatgctaccagaaag60
gcactgaagtcccagttccaggcagcagagtcatctacggcatgcaggacaccacgttgt120
<210>242
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>242
ttgtagctttgctgattcatgctaccagaaaggcactgaagtcccagttccaggcagcag60
agtcatctacggcatgcaggacaccacgttgtacaagaatgtaggttcattgtgctgctt120
<210>243
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>243
tttctaggaattctgtggcgaaaataacaacccacagtcatctgcacagcaattattatt60
cattataactgaactttccctctaaaaccgaagcttccctttcttactgcaaaacccaac120
<210>244
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>244
ctcaaaaggcattgtgaagcacatcagctgaggagtgagttataaatgaatctaacagag60
agacatttaatttccttctacataagcttgaggctgaagaggaaaagttagtgggcataa120
<210>245
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>245
gaaatggaaaatttaattaaagacaggaaactgaatttaatgatctctactgatctctta60
aattccccaaattatttcattatggcttacttattgaattttcttaaggaattctaaaat120
<210>246
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>246
aactgaatttaatgatctctactgatctcttaaattccccaaattatttcattatggctt60
acttattgaattttcttaaggaattctaaaataaatgaacaattacctgaattttgcagt120
<210>247
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>247
gcttctctctatgtatttcttgagagtttggacccatataactttagccgtcttttgaag60
attgttggccacaaaagagaaagcggacaactctagacctagggccacagtgtcaaggga120
<210>248
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>248
tcacctgagaggtgctctgggactttttcaatcctccctcctcacactccagttacctaa60
tttgggtcaggtagttctgaaaataaaatgctgaagttttcttttcctgcagaacaatgt120
<210>249
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>249
gggactaaccaactgctcttggaaaaacatggctgtgctaggaaattgacgttttcacgc60
cagtgggagtcgagcctgatgtgggtcaggaggcctcttcatctgttctgctggcttcag120
<210>250
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>250
agcacctaagggtagaggaaggaaaagggtttcctgttaccatattaagggctgagtctt60
tataccgcccattctccctctcagttcaggccagctccttgacctcagtttaatgaattt120
<210>251
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>251
ggatcatttctagacattgacaggccactgacaactacccccccaccccggaatctgagg60
ctgaggccgccccacaaaactgcattttgaaatcctttctacctctgggtatgggcaaag120
<210>252
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>252
cccagggattccgtgaggattcagccccatgcgaccatggcactccctgccacctgcctg60
cacaggccctgagcagagcctggctccagcccaaagggagacacacatccatgccattaa120
<210>253
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>253
tgtgttgcttgtttttgttgaaatgagagtattgcaaataatggattaggttgataccat60
ttgtttgttaatttgttttggatagtaaaccctgatttaggaaatgttcacataagaaat120
<210>254
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>254
agagactaaaacacgtcccagggttgcagaagaattaagttggagacaatgatgcccaac60
gatcaaaggggaaaataaaaagtaatcctgatgttaatatcagggagacagataagagct120
<210>255
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>255
ggaaacttgagggaatgtcctaactgcgaggtctattacatggcagaatagtctattaga60
tgcagaagcagtctccttggggatacagcagcaccatccctgtgggagtcatttaaaaca120
<210>256
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>256
actttgcccaccccacctcctctgtgcctccaacggtggccctgccacctgcacctgctt60
ttcctgcctgatatttataaacaagccatctccttcaccagctaagaaacagagaaagct120
<210>257
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>257
gctcactccaaactcgcggccgccctgccccgcctgtcatcccatagtacccaagattcc60
cactttacagcattccattttcttccaagaattcccctccaggcatcacactgtgcccct120
<210>258
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>258
cagacttggccggagcaaagccaactaaataaggtgcagctgggggcagaatgtgcttct60
ccaagcccaggccctcggccgcccagtgctgccccagtgcctttggctgcgacatctcag120
<210>259
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>259
gttcactgagttgggttcctaatctgtttctggcctgattcccgcatgagtattagggtg60
atcttaaagagtttgctcacgtaaacgcccgtgctgggccctgtgaagccagcatgttca120
<210>260
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>260
gcgagccccagccaatgagcgccggcgggccggttgcccaggcgaccagtgcgcggctcc60
gccccccgcggcgaggctcccgcgcgcggctgagtgcggactggagtgggaacccgggtc120
<210>261
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>261
cagcatgctaaccgccttcctccccagcatgctaaccgccttcctccccagcatgctaac60
cgccttcctccccggcatgctaaccgccttcctccccggcatgctaaccgccttcctccc120
<210>262
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>262
actgaggtggtggcattgggattggagaggagccagaaggcaggtagggcaggctttctt60
tcttctggaagccccagcaggcaggcatttgtgtccagataaggtgggctggccggaccc120
<210>263
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>263
ttaagtgaatttttcttttttcctgaactcctacagttgtgttttttttttgtgtgtgaa60
taaagtgatttttctttttttctgaactcccacagttgtgtttgtatgtgttggggagag120
<210>264
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>264
tgcgaaaggaaaagggagatgccttttcttgggattgctgggattaaaccagcttagaat60
ctggatttaagtataaaggccacatggctgctctctctgcaaggtttgaatttgctccaa120
<210>265
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>265
aattcagtgatgttcttacagttacaggtattctaaagaaactaatatcaattcatcaga60
aaattcaacatcgaccttatccacttgtttaattaatcaaattttatcactcaatagagc120
<210>266
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>266
ctcggccccgcgcagctccctccggcgtcccgcgccctccttccttcctcgccaggacgg60
acgcacttccggcggatgtgggggccagccctcggaaacggaagtgagcggcggggtcga120
<210>267
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>267
tcggaaacggaagtgagcggcggggtcgactgacggtaacggggcagagaggctgttcgc60
agagctgcggaagatggtgagtgggtacccgggcggctggagttccacgcgggctcggga120
<210>268
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>268
aataatgttttttatttgtgttgtagaatgccagaggacttggatctgagctaaaggaca60
gtattccagttactgaactttcagcaagtggaccttttgaaagtcatgatcttcttcgga120
<210>269
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>269
atatttaatctcctctaaatgttttttttttaatgtcctttcagtttttcttgtgtgaaa60
aatgaacttttgcctagtcatccccttgaattatcagaaaaaaatgtaagtatattatta120
<210>270
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>270
ttaaaattacattgtcttttgtagttccagctcaaccaagataaaatgaatttttccaca60
ctgagaaacattcagggtctatttgctccgctaaaattacagatggaattcaaggcagtg120
<210>271
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>271
tttcccctgagttttttctaatcttgtcccttcatacttttccccaaaggttcagcgtct60
tccatttctttcaagctcaaatctttcactggatgttttgaggggtaatgatgagactat120
<210>272
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>272
tttgagagtcttttaaaatgtttgtttttgtttgctttcagatccatcacaaagcgaagt60
catgggagagccacacttgatggtggaatataaacttggtttactgtaatagtgtgctgt120
<210>273
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>273
aattaagaagtataacaaaatcacatacatttgttccacggtttgaataaaatttccaat60
aactttcaaataatgcacacatagtatcgcatggatcaaattccgtacatttcaatatgg120
<210>274
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>274
aattaagaagtataacaaaatcacatacatttgttccacggtttgaataaaatttccaat60
aactttcaaataatgcacacatagtatcgcatggatcaaattccgtacatttcaatatgg120
<210>275
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>275
atgaaagttacaatatgcatgccatattgacacctattcaatagataaaatctaggaaaa60
tcttttttgtgaatactatttatactggaatgcaaattcacataaaaatacttcctttaa120
<210>276
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>276
ataaaatctaggaaaatcttttttgtgaatactatttatactggaatgcaaattcacata60
aaaatacttcctttaaagcatgttctatttcacacaagaaattaaatgtgagctactttt120
<210>277
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>277
gttctatttcacacaagaaattaaatgtgagctacttttaaaattgcaagctttattttt60
aatcaaaccaattcaattttaagtttttagatcattaagtctcatacaacttcctctaat120
<210>278
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>278
catcgacagggttttattgttacatttggccttaaccttaaaaaacaaatttgtttacat60
ttcatatttaaatgggacattttaaaccctgagtaaaaagacatatttttacaagtccag120
<210>279
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>279
ttttattgttacatttggccttaaccttaaaaaacaaatttgtttacatttcatatttaa60
atgggacattttaaaccctgagtaaaaagacatatttttacaagtccagctgtaacaatt120
<210>280
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>280
tgcagtgcttttattagataaaaactattgttaacgttttgactttttagaatatactgt60
ttagaaggttaatgctttttcatgactttaaaagtttctaaatgtattttattagtatct120
<210>281
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>281
tgtttagaaggttaatgctttttcatgactttaaaagtttctaaatgtattttattagta60
tctaagtaaaaattctagatttctatatggatattataaaaacacctcttctttagactg120
<210>282
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>282
tctatatggatattataaaaacacctcttctttagactgtttatttcaccttgatcataa60
aaaatcattgaatactacattcaagataatttttctttcttggctttgggctattgtctt120
<210>283
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>283
ccttgatcataaaaaatcattgaatactacattcaagataatttttctttcttggctttg60
ggctattgtcttttcaaccttaaaagttaaaattattgttttcctatatgtaatgatact120
<210>284
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>284
tgtctgattttttctgcacgtcctttacttttctgtattcgttctaaagttgaacacaaa60
tctgcaaaaggaacgtgagatgcaacagtttgtgaagtttgtgaggcagccatcaaagcg120
<210>285
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>285
gcttgggtctagatgaagagcacagcatcaggaggccaggcttctaatgcggcgcagtct60
acagcgctgtggactcggcaactcccatcacctgccgctgcacaaacaccctgcttgctg120
<210>286
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>286
cccagcctcagtctctccaactgagttcctgcctgcctgcctttgctcagactgtttgcc60
ccttactgctcttctaggcctcattctaagccccttctccaagttgcctctccttatttc120
<210>287
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>287
attgtgccggcacatgaatgcaccaggtgttgaagtggaggaattaaaaagtcagatgag60
gggtgtgcccagaggaagcaccattctagttgggggagcccatctgtcagctgggaaaag120
<210>288
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>288
gtcttgcttttaagtccaaagcctgagcccaccaaactctcctacttcttcctgttacaa60
attcctcttgtgcaataataatggcctgaaacgctgtaaaatatcctcatttcagccgcc120
<210>289
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>289
acagctaatgagtggaggaagagagacacttgtgtacaccacatgccttgtgttgtactt60
ctctcaccgtgtaacctcctcatgtcctctctccccagtacggctctcttagctcagtag120
<210>290
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>290
tcctcattccagatctgcaagattgtaagacagcctgtgctccctcgctccttcctctgc60
attgcccctcttctccctctccaaacagagggaactctcctacccccaaggaggtgaaag120
<210>291
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>291
aaaaaggttattggttggcagttcagccaatgagcgggcctgttactagtttgcggcatc60
ctgtggtataggggaagcgctccgggcctggaatccctacgcgtccctttgggtttagca120
<210>292
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>292
ttaagtatgacctgtcagcctctacattctctcctgacggaagagtttttcaagttgaat60
atgctatgaaggctgtggaaaatagtaggtaagaaacatttaaccaggtattacatgtct120
<210>293
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>293
ctccttgtttcagtacagctattggaatcagatgcaaagatggtgttgtctttggggtag60
aaaaattagtcctttctaaactttatgaagaaggttccaacaaaagactttttaatgttg120
<210>294
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>294
tagcaggtttgttggcagatgctcgttctttagcagacatagcaagagaagaagcttcca60
acttcagatctaactttggctacaacattccactaaaagtaagttgataacatattgagg120
<210>295
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>295
aataagctcttctgcttccctccatagcatcttgcagacagagtggccatgtatgtgcat60
gcatatacactctacagtgctgttagaccttttggctgcaggtaagaaattttgtgaatt120
<210>296
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>296
attgccatggtttaagctgtttgtatactttcatgcagtttcatgttagggtcttacagt60
gtgaatgacggtgcgcaactctacatgattgacccatcaggtgtttcatacgtgagtaat120
<210>297
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>297
ttttttctaaatagggttattggggctgtgccatcggcaaagccaggcaagctgcaaaga60
cggaaatagagaagcttcaggtaataattaatgcttgaatttttactatgtcgtcatcct120
<210>298
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>298
acaagtggaaaatgttcttctgatgaacagtatacattttattttctatagatgaaagaa60
atgacctgccgtgatatcgttaaagaagttgcaaaaatgtaagttgaaatttttcttacc120
<210>299
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>299
tggaggttaccaaaatcttacctttcccttttctattctagaatttacatagtacatgac60
gaagttaaggataaagcttttgaactagaactcagctgggttggtgaatgtaagttattt120
<210>300
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>300
tactcaatttaaaattcatttacacatgtggcttaatttacagtaactaatggaagacat60
gaaattgttccaaaagatataagagaagaagcagagaaatatgctaaggtaagccacagc120
<210>301
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>301
atgatgataatatgtaacatttactccagcatctattgtattttaaatttctactccagt60
ccaatgtaactatttagccctggattatacatactgtccaattttcattaaatttttgtc120
<210>302
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>302
cagtagcaggtgccatccgcctccgccatgacaacagacacattgatgtgggtgggttta60
cccgccatgcggtcgatggtcttctgtgtgaaggccagcggcagggcctcgtggcccacc120
<210>303
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>303
tcctgtgacccctgcagccagcgaaccagcacatccttggggctgaagccacgtgccagg60
cacgtcagcgtcaccagctcgttcagggccagctcctccgacggcggcggcagcaggtgg120
<210>304
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>304
ggattttgtgatgttggcggttagtggggtcttcaactcggggtgggcagcagtgcaggt60
gaaggtctccccatggttccatggctgggcacagccaggcaggacactggacacgctgta120
<210>305
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>305
tccgctttcgctccaggtcacactgagtggctcctgggggaagaagccctggaccaggca60
tgcgacgaccacgttcccatcttggggggtgctgtcgaggctcagcgggaagaccttggg120
<210>306
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>306
cctcgttgtgcagccactgcaccgagatgtcctcaggcatgaagttctggatcaggcagg60
cgagggtgcgcttgtcccggctccccggccactccggcgtcgcaaacgcatagacttccg120
<210>307
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>307
aggttcacggtccccttgctgggtgccaggtccaccaccagacaggtgatcgtgggcgac60
ttgcggatgaacaggtcgaacgggctgggccggcttaggtaggcgctcacccctctcggg120
<210>308
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>308
atagtccctggggtgtacccagagacgaggcacaggagctggatggtcggggggaagtgc60
ccgccgccgtcgcaggacgactgtaagatcttcacggtgggcggggtgaagtccctggag120
<210>309
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>309
ggagcctgtgtcccaggtcaccatcaccggctccgggaagtagcccgtggccaggcagcc60
cagagtcacggaggtggcattggagggaatgtttttgcagcagcgggtcaaggggaagac120
<210>310
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>310
tctcccactccacggcgatgtcgctggggtagaagcctttgaccaggcaggtcaggctga60
cctggttcttggtcatctcctcctgggatgggggcagggtgtacacctgtggctctcggg120
<210>311
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>311
aactggacctcggggtcttcctggctcacgtccaccaccacgcacgtgacctcaggggtc60
cgggagatcatgagagtgtccttgggttttggggggaacaggaagactgatggtcccccc120
<210>312
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>312
ctggagggcgaggcctgggttggcttacctgggcatgatgggcatgggggaccatatttg60
gactctgcagagagaagattgggagttactcagatctgggaggagagaaggtgtctgagc120
<210>313
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>313
aactctcttgtccaccttggtgttgctgggcttgtgatctacgttgcaggtgtaggtctt60
cgtgcccaagctgctggagggcacggtcaccacgctgctgagggagtagagtcctgagga120
<210>314
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>314
catttacccggagacagggagaggctcttctgtgtgtagtggttgtgcagagcctcatgc60
atcacggagcatgagaagacgttcccctgctgccacctgctcttgtccacggtgagcttg120
<210>315
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>315
aactggacctcggggtcttcgtggctcacgtccaccaccacgcacgtgacctcaggggtc60
cgggagatcatgagggtgtccttgggttttggggggaagaggaagactgacggtcctgcc120
<210>316
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>316
tactctagggcacctgtcccgccttgagctggagggcgaggcctgggctggcttacctgg60
gcacggtgggcactcgacacaacatttgcgctctgcagagagaagattgggagttactcg120
<210>317
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>317
aactgtcttgtccaccttggtgttgctgggcttgtgatctacgttgcaggtgtaggtctg60
ggtgccgaagttgctggagggcacggtcaccacgctgctgagggagtagagtcctgagga120
<210>318
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>318
gttccacgacaccgtcaccggttcggggaagtagtccttgaccaggcagcccagggccgc60
tgtgctctcggaggtgctcctggagcagggcgccagggggaagaccgatgggcccttggt120
<210>319
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>319
taccccggggacagggagaggctcttctgcgtgtagtggttctgcagaccctcatgcatc60
acggagcatgagaagacgttcccctgctgccacctgctcttgtccacggtgagcttgcta120
<210>320
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>320
acttgacctcagggtcttcgtggctcacgtccaccaccacgcacgtgacctcaggggtcc60
gggagatcatgagggtatccttgggttttggggggaagaggaagactgacggtcccccca120
<210>321
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>321
ggcacctgtcccgccttgagctggagggcgaggcctgggctggcttacttgcacatggtg60
ggcatgtgtgagttgtgtcacaacatggggttttgggctctgcagagagaagattgggag120
<210>322
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>322
tcttgtctatccgcgagggtccaggcagggtccagtgtctgggctcacgggcattgagtg60
tgcacctggctggtgccacctgcctcaccttagccccctccctgccccaaagccaaagtc120
<210>323
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>323
cagtagcaggtgccgtccacctccgccatgacaacagacacattgacatgggtgggttta60
cccgccaagcggtcgatggtcttctgtgtgaaggccagcggcagggcctcgtggcccacc120
<210>324
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>324
tgcacgtgaggttcgcttctgaacctaagagcaggtcctcgagggccggtcggtgcagtg60
acagtcgggggtggcagcatgagggagatggggtaggtggagttgagggagatggggtag120
<210>325
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>325
tgggcagggcacagtcacatcctggctgggattcgtgtagtgcttcacgtggcatgtcac60
ggacttgccggctaggcactgtgtggccggcagggtcagctggctgctcgtggtgtacag120
<210>326
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>326
cctcgttgtgcagccactgcaccgagatgtcctcaggcatgaagttctggatcaggcagg60
tgagggtgcgcttgtcccggctccccagccactccggcgtcgcaaacgcatagacttccg120
<210>327
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>327
gctggtcttggtcgtggaccgcacgagggccctgggcaggtgggggtgggtcaccctgca60
ctggtaggtctccccttcgatccagtctcgggtgcccaccggcagggtggacgtgacggt120
<210>328
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>328
tctgtgtgtagtggttgtgcagagcctcatgcatcacggagcatgagaagacgttcccct60
gctgccacctgctcttgtccacggtgagcttgctgtagaggaagaaggagccgtcggagt120
<210>329
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>329
aacttgacctcagggtcttcgtggctcacgtccaccaccacgcatgtgacctcaggggtc60
cgggagatcatgagggtgtccttgggttttggggggaagaggaagactgacggtcccccc120
<210>330
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>330
ctagggcacctgtcccgccttgagctggagggcgaggcctgggctggcttacctgggcac60
ggtgggcatgtgtgagttttgtcacaagatttgggctctgcagagagaagattgggagtt120
<210>331
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>331
aactttcttgtccaccttggtgttgctgggcttgtgattcacgttgcagatgtaggtctg60
ggtgcccaagctgctggagggcacggtcaccacgctgctgagggagtagagtcctgagga120
<210>332
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>332
ggccctaggccccctgcccaatcatgttcctatagtcggggatgatggtctgcttcaggt60
ccaccaccgaggagaagatccacttcacctgtaggcaaggcacagcacaggggtgagcga120
<210>333
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>333
gcggccgaccttgaagaaggtgacggtggcactgtagcacacgcttaacaggaagagtgt60
gatgaagatggtgatggtcgtccacagcccgtccagctccccgtcctgcgcctccgcaca120
<210>334
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>334
tctcccactccacggcgatgtcgctggggtagaagcctttgaccaggcaggtcaggctga60
cctggttcttggtcatctcctcccgggatgggggcagggtgtacacctgtggttctcggg120
<210>335
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>335
aactggacctcggggtcttcgtggctcacgtccaccaccacgcacgtgacctcaggggtc60
cgggaaatcataagggtatccttgggttttggggggaagaggaagactgacggtcctccc120
<210>336
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>336
ctagggcacctgtcctgccttgagctggagggcgaggcctgggctggcttacctgggcac60
cttgggcacgggggaggtgtgtcacaagatttgggctctgcagagagaagattgggagtt120
<210>337
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>337
ctagggctcttgtcccgccttgagctggagggcgaggcctgggctggcttacctgggcac60
cgtgggcatgggggaggtgtgtcacaagatttgggctctgcagagagaagattgggagtt120
<210>338
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>338
ctagggctcttgtcctgccttgagctggagggcgaggcctgggctggcttacctgggcac60
cgtgggcacgggggaggtgtgtcacaagatttgggctctgcagagagaagattgggagtt120
<210>339
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>339
gggctcttgtcccgccttgagctggagggcgaggcctgggctggcttacctgggcaccgt60
gggcatgtgtgagttgtgtcaccaagtggggttttgagctctgcagagagaagattggga120
<210>340
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>340
aactctcttgtccaccttggtgttgctgggcttgtgattcacgttgcaggtgtaggtctg60
ggtgcccaagctgctggagggcacggtcaccacgctgctgagggagtagagtcctgagga120
<210>341
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>341
tcacagacttcctagagttcattcaacaagagtttattgtggttgtactgtgagctcggc60
cagagctgctgagtggcgggtgtggacggatctggtccgggatcatttcatggggccatg120
<210>342
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>342
gctgggcggggctgggacacgcagcacactccaggcccagaacgtggtgctcctgggctg60
tggaggggggcgtgcgggggcaaacccagaagtgttcacctcacgctggtcctccagcca120
<210>343
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>343
gggttctctcagcgccatcaacctctggggtgggaggctgggatggttcagtgtgcaggt60
gacggaggtccccgcgttccacaaggacctgggcagggtcagacggctgtgctggctctg120
<210>344
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>344
agtttagggcaccaacggccaactagttattgggcaataggtgttttgatagcccagggg60
tcagaaaaaacctggtgtctttgtctctctctcttcttgttcctctttctccttctcctt120
<210>345
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>345
tgtgcagggaaggggcttctcacctgtgttacgggtggtggctggggctgtggttgcctt60
ggcgaggctgccctctgcttggggttgtgcagtgggcactgaggaggcctgtgcctttgg120
<210>346
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>346
tggccagcggaagatctccttcttactcttgctggcggtgtgctggaccacgcatttgta60
ctcgccttggcgccactgctggaggggggtggagagctggctgcttgtcatgtagtagct120
<210>347
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>347
ccctttgagttcctctctctggtctccgatgttcttctgttgggatcatttcacctacag60
gcaacagagacagtgtgaaatgctttccccgtggtcgggaagggagccggggcagagatg120
<210>348
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>348
gccacagccgtgctaccttgaacaaggtgacggtggtactgtagaagaggctcaggagga60
agaggacgatgaaggtggaggcggtggcccacaggttctcaaagccctcctcgtcggcgc120
<210>349
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>349
acctccaagcagagacagaaggacaagcagagcctgggcccaaagccccggtgccagcac60
agcgcagggagcaaactgctgcagggacagagtatccggcagggccctgggagaatgttg120
<210>350
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>350
tccacacatgcctgtgtgctgcctgtatacgtgtggccttgctttcagctttctttctgg60
aagcagtaggaggaatggcaagtctgaggtcagcctgctgtgagcagctgcagggaattc120
<210>351
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>351
cccacaaaaacccagccacccagagaccttctgtctccgggcgtcacatggaagctgact60
gtccgtggccctgtcctgcccttctcatggaaccctctgctggcctcccacgtaccccac120
<210>352
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>352
ccacgtaccccacattctggcctgacccctcagaagccagaccactgtcggcctgggaag60
tccaactgcaagcagacggctgctaagtcacccccaggagtccaaaaaccccggggggca120
<210>353
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>353
agagggggtggtgaggactcacctgaggagacggtgaccagggttccctggccccagggg60
tcgaaccagttgtcacattgtgacaacaatgccaggaccccaggcaagaactggcacccc120
<210>354
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>354
tgcagcaaaacccttcagagttaaagcaggagagaggttgtgaggactcacctgaggaga60
cggtgaccagggttccctggccccagtagtcaaagtagtcacattgtgggaggccccatt120
<210>355
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>355
ggcagaaggaaagccatcttacctgaagagacggtgaccattgtcccttggccccagata60
tcaaaagcatcacacagggacacagtccctgttcctgcccagacataaacctgtgcccgt120
<210>356
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>356
cagccacacagacccccgggctgagacccaggcagggaggggtgacgttcccagggagac60
ggtggccgggctgccctggccccagtgctccaagcacttgtagccacactaaagcgcagg120
<210>357
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>357
gggggctgcagtgggactcacctgaggagacagtgaccagggtgccacggccccagagat60
cgaagtaccagtagcacagcctctgccctcctgcttctcccatacaaaaacacaccctcc120
<210>358
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>358
cctggcccagtacacctggctccccgctatccccagacagcagactcacctgaggagacg60
gtgaccagggtgccctggccccagtgctggaagtattcagccacggtgagtcagccctga120
<210>359
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>359
acagtggttctcagctcagccaaaaccctgcggctggtagggggcctgtggggctggggg60
ctgatgtggctgcggtctccagggcgggtcgggtccaggggcccccagcacctttgccag120
<210>360
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>360
cttctctggctgggcctccagcagtggccgggagacagttttggacatagcttttctcac60
agtggtagtagctcccactatacccaccgtgactcggggctgttcagaatccacccaggc120
<210>361
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>361
ttgggatggggtttctgtcctgcccagtgtcattgtggtagccgctgctatacccgactg60
tgacacagacagcttcagaaacctcctggtttgtttgggggccttcccttgctgggctca120
<210>362
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>362
ctggggctgcgtggcaggggtctgtctacttggcctggttgctgctatgggcggcaccac60
tgtggtaattgtagccatctctaccacggcctggcaccccctgacaataaccacacctgg120
<210>363
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>363
gagccgagaagtgtgcataaaccctgcgagtccggtcagtttttgctgctggtttcatca60
ctgtgggagttaccaccgtagtcacacagcaggagggcccttcacaaaaagcccctgagt120
<210>364
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>364
atgagtttttgatggactctgtgacactgtggtagtaataaccactactatcatagtaat60
accacagtgacacagacctcacttcaaacctaccgccaggcctggggaaacccgggatgt120
<210>365
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>365
tcagtcccaggagtgggtgcgtttgcagtagggctttaggaatggggttgtgtcactgtg60
ggaatagcagtcaccaccacaatatgctcacagtgacacgaacccccacaaaatcctcct120
<210>366
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>366
gccaggagtcagttttggacacagtttttctcacggtggtcgttccagttatacccactg60
tgactcggggctgttcagaatccgaccaggcgccctgagctctggggcctcctgggtggg120
<210>367
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>367
ctccctgctctctctgcttccgctgacttgggacggggtttctggcctggcgagtgtcac60
tgtggtaccagccactgctataccccactgtgatacagacagcttcagaaacctcctcgt120
<210>368
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>368
ctggggctgcacagcaggggcctgtctacttggcctggttgctgctatgggcagcaccac60
tgtggtaaccatagctgtatccaccacagtctgacaccccctgacaataaccacacctgg120
<210>369
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>369
gtgtccggccagtttttgctgctagtttcatcactgtggtagtcaccgtagtcacacagt60
aggaggggccttcacaaaaagcccctgagtgtgcccagcggcctttcccacgcaggccct120
<210>370
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>370
ggtcccctgcggggagggctgtggcatgggtttctgatggaccgtgtgatgctgtgggta60
taacgataactcccccaaacgtaatcataataccacagtgacacagacctcacttcaaac120
<210>371
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>371
cagccccaggagtgggtgtgtttgcagtagggctttgggaatgggtctgtgtcactgtgg60
gagtagcagctaccaccactacaatatcctcacagtgacacgagcccccacaaaatcctc120
<210>372
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>372
gccactggacagtttttgacgtagctattctgacagtggtggttccggttatacccgctg60
tgactcggggctgttcggaatccgacggggcgccctgagctctggggcctcctgggtggg120
<210>373
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>373
ctccctgctctctctgcttccgctgacttggaatggggtttctggctgggtgagtgtcac60
tgtggtaccagctgctgctataccccactgtgacacagacaccttcagaaacctcctggt120
<210>374
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>374
ttggcctggttgctgctatgggcggcaccactgtggtaatcgtagccactatatccacca60
cagtctgacatcgcctgacaataaccacacctggaactggaggcggggctgtcaggagga120
<210>375
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>375
gtttccagccagtttttgccactgggttcatcactatggtagttactgtagtcacacagc60
aggagggcccttcacaaaaagcccgagtgtggccagagacctttcccacgcaggccctgg120
<210>376
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>376
atgggtttttgatggactctgtgacactgtggttataataactccccgaaccatagtaat60
accacagtgacacagacctcaccccaaacctaccgcccagcctggggaaacccgggatgt120
<210>377
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>377
atgggtttttgatggactctgtgacactgtggttataataaccagtcaaaatatcgtaat60
accacagtgacacagacctcattctaaacctaccggcaggcctggggaaacctgggatgt120
<210>378
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>378
agccccagaagtggaatgcgtttgcagtagggctttgggaatggggctgtgtcactgtgg60
gtatagcatacaccattagtacaatatcctcacagtgacacgagcccccacaaaatcctc120
<210>379
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>379
gccaggagaccgttttggacgaagcttttctcacagtggtagttccagttatacccactg60
tgactcggggctgttcagaatccgacggggcgccctgtgctctggggcctcctgggtggg120
<210>380
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>380
ttgggatggggtttctggcctggcgagtgtcactgtgggacgagctgctatactccactg60
tgacacagacaccttcagaaacttcctggtttgtttggggaccttcccttgctgggctca120
<210>381
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>381
ctggggctgcacagcaggggcctgtctacttggcctggttgctgctatgggcagcaccac60
tgtggtaaccatagctgtatccaccacagtctgacaccccctgacaataaccacacctgg120
<210>382
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>382
tgagcggagaagtgtgcataaaccttgggagtccggtcagtttttgctgctgggttcatc60
actgtggtagttactgtagtcacacagtaggaggacccttcacaaaaagcccctgagtgt120
<210>383
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>383
atgggtttttgatggactctgtgacactgtgggtataataaccactccaaaaatcgtaat60
accacagtgacacagacctcaccccaaacctaccgccaggcctggggaaacccgggatgt120
<210>384
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>384
cagctccagaagtggaagcgtttacagcagggctttgggaatggggctgtgtcactgtgg60
gcatagcagctggtactactacaatatcctcacagtgacacgagcccccacaaaatcctc120
<210>385
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>385
cccctctctggcgcggcctccagcaggggccaggagagagttttggacacagtttttctc60
acggtggtcgttccagttgtacccaccgtgactcggggctgttcagaatctgccccggcg120
<210>386
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>386
ctcttgcacagtaatacacagccgtgtcctcgggagtcacagagttcagctgcagggaga60
actggttcttggatgtgtctgggttgatggttattcgacttttcacagatactgcataat120
<210>387
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>387
acggcatctccctgacactgaccccatgggaggcccagcacgggcaggaagatgaggaag60
gagacagacattgtctggagctctggtgactgccctggtcagccagaattcagcaggctc120
<210>388
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>388
caatatctaaattagctagcaaaactctacagggtgattaaaaaaaaaaaaaaaaaacaa60
ctgtgaaggtgagaggtgcagtgagtccttggcagaggccgtttgctcagggaaagtaaa120
<210>389
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>389
ctctcgcacagtaatacacggccgtgtcgtcagatctcagcctgctcagctccatgtagg60
ctgtgctgatggacgtgtccctggtcatggtgacccagccctgaaacttctgtgcatagt120
<210>390
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>390
ctggactggactcaatctctttctcatcactgggactagggagcctcttacctgtggctg60
ctgccaccaagaagaggatcctccaggtccagtccatggtgaggaacggagctctcaggg120
<210>391
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>391
cactcacagtaatacatgaccttgtccttggctttcagattggtcattgtaaggcagact60
gcacttaaaagggtgttgcttgagattgttaatttatttgtactcatggagagtaaccct120
<210>392
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>392
ctctcgcacagtaatacacagccgtgtcttcagatctcaggctgctcagctccatgtagg60
ctgtgctcgcggatgtgtccctggtaatggtgactctgccctggaacttctgtgaatatt120
<210>393
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>393
cttgactggcctcgatcccttcctcctcactgggatttggcagccccttacctgtggctg60
ctgccaccaaaaagaggatcctccaggtccagtccatggtgaggcgttgtgctctggggg120
<210>394
<211>120
<212>dna
<213>人工序列(artificialsequence)
<400>394
ggatcagaggcacctcccatacccccatgtctgcagccttgagacactcacatctgggag60
ctgccaccaggaggaggaagaaccacaggtgtttcatgttcttgtgcaggaggtccatga120
<210>395
<211>15
<212>dna
<213>人工序列(artificialsequence)
<400>395
gactacatgggacat15
<210>396
<211>15
<212>dna
<213>人工序列(artificialsequence)
<400>396
ggaacctacgacgta15
<210>397
<211>50
<212>dna
<213>人工序列(artificialsequence)
<400>397
aatgatacggcgaccaccgagatctacactctttccctacacgacgatct50
<210>398
<211>50
<212>dna
<213>人工序列(artificialsequence)
<400>398
caagcagaagacggcatacgagatcggtctcggcattcctgctgaaccgc50
<210>399
<211>21
<212>dna
<213>人工序列(artificialsequence)
<400>399
aatgatacggcgaccaccgag21
<210>400
<211>21
<212>dna
<213>人工序列(artificialsequence)
<400>400
caagcagaagacggcatacga21