本发明属于病毒分子检测
技术领域：
，具体涉及一种鲤冠状病毒hl39的rt-lamp检测引物及应用。
背景技术：
：套式病毒目(nidovirales)于1996年首次由国际病毒分类委员会(internationalcommitteeontaxonomyofviruses,ictv)提出，该目病毒宿主广泛，可引起哺乳动物、鸟类、爬行动物、鱼虾等水生动物多种重大疾病。套式病毒目冠状病毒科是一类线形单股正链rna病毒，最早于1937年从鸡身上分离出来，almeida等对这些病毒进行了形态学研究，电子显微镜观察发现这些病毒的包膜上有形似日冕的棘突，故提出命名这类病毒为冠状病毒[1]。1975年，国际病毒命名和分类委员会正式命名了冠状病毒科。由冠状病毒科的严重急性呼吸综合征病毒(sars-cov)引起的非典型性肺炎，世界上有30多个国家和地区相继报道出现病例，病死率超过10％。2012年，中东地区开始出现新型冠状病毒，病死率超过50％，引起了世界范围的广泛关注，造成社会对冠状病毒的恐慌。因此，对冠状病毒进行深入研究具有重要意义。2018年ictv调整了套式病毒目的分类，新增了abyssoviridae、medioniviridae、mononiviridae、euroniviridae和tobaniviridae等5个病毒科，其中在tobaniviridae科下新增oncotshavirus属。2017年，申请人发表了鲫冠状病毒的相关研究成果，目前鲤冠状病毒(cyprinidnidovirus,cynv)尚无公开报道，更也无有效的检测方法。申请人首次从鲤鱼病变组织中分离出一株鲤冠状病毒，针对该病毒，设计了rt-lamp引物，从而实现鲤冠状病毒的快速、精确检测。技术实现要素：本发明的目的在于提供了一种鲤冠状病毒hl39的rt-lamp检测引物，所述的引物为引物组：f3：5’-tgcccaccaagacgacta-3’；b3：5’-tttgcgctagcgcgattc-3’；fip：5’-cgttccaaaccgtcttgggtcaagagtgacgtaactgctactg-3’和bip：5’-actcacttctgctccttgcgacgcagagtgtaagaggttttcca-3’，所述病毒的保藏编号为cctccno：v201988。本发明的另一个目的在于提供了一种鲤冠状病毒hl39的rt-lamp检测引物的应用，利用该引物，可制备成鲤冠状病毒hl39的检测试剂盒。为了达到上述目的，本发明采取以下技术措施：一种鲤冠状病毒hl39的rt-lamp检测引物，所述的引物为引物组：f3：5’-tgcccaccaagacgacta-3’；b3：5’-tttgcgctagcgcgattc-3’；fip：5’-cgttccaaaccgtcttgggtcaagagtgacgtaactgctactg-3’和bip：5’-actcacttctgctccttgcgacgcagagtgtaagaggttttcca-3’，所述病毒的保藏编号为cctccno：v201988。鲤冠状病毒hl39的rt-lamp检测引物在制备成鲤冠状病毒hl39的检测试剂盒中的应用；以上所述的应用中，优选的，所述的鲤冠状病毒hl39的检测试剂盒包括：fip、bip、f3和b3，dntps，betaine，mgso4，bstdna聚合酶，amv逆转录酶，模板rna，10×thermopolreactionbuffer。与现有技术相比，本发明具有以下优点：本发明公开了一种鲤冠状病毒hl39的rt-lamp检测引物，利用该引物制备的试剂盒，特异性好，能有效检测出cynv病毒；灵敏性好，最低检测限可达12.27pg/μl的rna；快速高效，检测时间约1小时，非常适用于cynv-hl39的现场快速检测。利用本发明的引物制备的rt-lamp检测试剂盒不需要特殊试剂与设备，检测成本低。结果判定简单，通过反应过程中大量产生的焦磷酸根和体系中的镁离子形成白色稳定的焦磷酸镁沉淀，经肉眼就可判断结果，或通过加入1000×sybrgreeni，可提高结果判定的灵敏度。具体实施方式本发明所述技术方案，如未特别说明，均为本领域的常规方案，所用试剂或原料，如未特别说明，均购自商业渠道或是已公开。实施例1：一种鲤冠状病毒hl39，其分离过程如下：1)将携带病毒的鲤鱼肝、脑、脾、肾等组织取出并碾磨后，培养基稀释100倍，2000rpm离心后取上清过滤，接种到草鱼性腺细胞(grasscarpovarycellline,gco)单层，25℃培养7天，产生细胞病变效应(cpe)，可见gco细胞收缩变圆、折光度增加，部分细胞开始脱落，细胞单层开始破裂，培养基：m199，10％胎牛血清，ph7.0～7.2；2)收集病变细胞，利用透射电镜进行形态学鉴定：鲤冠状病毒hl39的形态特征：病毒呈棒状，四周有明显的日冕状棘突结构，与典型的冠状病毒形态相符。3)收集病毒，对其进行全基因组测序，序列为seqidno.1所示。鲤冠状病毒的形态特征：病毒呈棒状，四周有明显的日冕状棘突结构，与典型的冠状病毒形态相符，该病毒已于2019年11月26日送至中国典型培养物保藏中心进行保藏，分类命名：鲤冠状病毒(cyprinidnidovirus)hl39，保藏编号：cctccno：v201988，地址：中国武汉武汉大学，本发明或称该病毒为cynv-hl39。实施例2：基于鲤冠状病毒hl39的全序列(seqidno.1所示)与其他病毒的全序列的差异设计的rt-lamp检测引物为：f3：5’-tgcccaccaagacgacta-3’；b3：5’-tttgcgctagcgcgattc-3’；fip：5’-cgttccaaaccgtcttgggtcaagagtgacgtaactgctactg-3’；bip：5’-actcacttctgctccttgcgacgcagagtgtaagaggttttcca-3’。实施例3：利用鲤冠状病毒hl39的rt-lamp引物对cynv-hl39进行检测，方法步骤如下：1)、病毒rna的获取：采用或ls试剂或柱吸附洗脱法提取样本总rna。2)、rt-lamp扩增：采用25μl反应体系：内引物fip和bip各0.8μm，外引物f3和b3各0.1μm，dntps1mm，betaine0.5m，mgso45mm，bstdna聚合酶8u，amv逆转录酶5u，模板rna5μl，10×thermopolreactionbuffer2.5μl，选择64℃和60min进行rt-lamp反应之后，80℃灭活2min。3)、检测结果判定，采用下述三种方式之一：a、发生扩增反应时，从dntps析出的焦磷酸根离子和反应液中的镁离子形成白色焦磷酸镁沉淀，通过肉眼判断：检测管出现明显浑浊为阳性，未见浑浊为阴性。b、每反应管加1000×sybrgreeni2μl，轻轻混匀，1-5min后在黑色背景下观察结果，反应液变绿为阳性，保持橙色为阴性。c、取扩增产物，用2％w/v的琼脂糖凝胶电泳后，置于凝胶成像系统中成像，电泳图片显示lamp特征性梯状条带，则结果为阳性；如无任何条带，则结果为阴性。实施例4：鲤冠状病毒hl39rt-lamp检测试剂盒特异性试验利用实施例3所述的方法，对表1中的待检病毒样品进行检测，同时设置阴性对照组(双蒸水)和阳性对照组(cynv-hl39)，特异性试验组具体为：鲤春病毒血症病毒(svcv)，草鱼呼肠孤病毒873(gcrv-873)，草鱼呼肠孤病毒873(gcrv-9014)，传染性造血器官坏死病毒(ihnv)。表1鲤冠状病毒rt-lamp检测试剂盒特异性试验试验编号名称扩增结果1cynv-hl39+2svcv－3gcrv-873－4gcrv-9014－5ihnv－结果如表1所示，仅阳性对照组(cynv-hl39)显示lamp特征性梯状条带，其余组均无特征条带，说明鲫鱼冠状病毒hl39rt-lamp检测引物具备很强的特异性。实施例5：鲤冠状病毒hl39rt-lamp检测试剂盒的灵敏度将1227ng/μl的cynv-hl39rna模板进行10倍梯度稀释，利用实施例2所述的方法进行检测，检测本发明提供的试剂盒的检测限，结果显示105倍稀释样品还可观察到特征性梯状条带，本发明提供的试剂盒最低可检测12.27pg/μl的病毒rna模板。序列表<110>武汉海关技术中心<120>一种鲤冠状病毒hl39的rt-lamp检测引物及应用<160>5<170>siposequencelisting1.0<210>1<211>25833<212>dna<213>人工序列(artificialsequence)<400>1ttgaattaccaataaactaccacccgctattatagcactagacactagcgaactgctagc60ttcctccaagctgcacaacgcaaactgtctttttaactctcctaacctcgaacgcagcgc120aggtaccctcacccggaccacgcgtgtgcccttgactggaaaccctaccttcgatcaggc180cttcgaacgcagcaaaggtaccctcacccggaccacttgtgtgcccctacgcctgctctt240acgtagccccatcctacccacccgccaactcttaacctatcagctttgcccaccgtgacg300actagacgtgatctaactgctactaccaatcctattacaaccaacttgcccaccaagacg360actagagagtgacgtaactgctactgcctaaggcactaccagcgaaatcataatctgctc420ctcttgacccaagacggtttggaacggtgctactagtgaactcacttctgctccttgcga480cccaaagacggttattatttggaaaacctcttacactctgcccctttacctactcaatat540aacttaaatctcttttagaatcgcgctagcgcaaaccttcaccggtaaccgccagaacgt600cctacaagctctcaaccaccggcactttgttgctaaatcctacgtgcaagcatctttttg660ctataacatccacgttcgaaaatcaacaaaaatgcccaacatgcgaatccgcaagattgc720tcctaaagccgctgttaaaccaacccatagcggcccaagagtggtctccaatcccgcaag780ctgccacatctacatcggaccccaactgcgtgctcacaactacccgagcgtgctaaaggg840ctcctttaaggctgtcagcgcggaaaccataacagctaccaacctaattgccaccgtccg900caaggatgctggtatttcaggtcaatatgctgggaagatgggttatgaccccgagatggg960acttttcgtctccgtcgaaggtgttcgtgctctccaagcttccaaactccaaactgccct1020taaggcaattaaaaagggtactagaccctttggccttttcaggcctaaacagagtgtgac1080taacacccacgctcaaaatttgctagccctccaccgtgctagctccaccggcccttggac1140caaacacttaccatctcaaggtcccataggtaactgcctcgcagaattcatgcaggagta1200cactaccacaattccaccatttgatagtgtcctcactgaaaacggttggcaatacgctgt1260cacgattgacggtgtgactaaatctcgcctcccgtacaccgaaattcgccgcggtggtta1320tgaatttaaaacaactaattaccacatcggtgcaatccaggttactgccaaccacattgg1380caaagtaaaccctggagatactaaattggactgtcaatatttcatgttggcacgtaatca1440aaagccccaaaacttcactgctgagcaactcgaaggaaacatccataacatcatcttcga1500cctcagtcatgaaactggcaaacccaaactctatggaatggaagaaacacgacaatgggc1560tgggacaacaccaaaaccacctaagaaaaagcttagacttgtagctatgacatcaactct1620cagtgccactccatcaaccacacacccagcgcccctaacaaccacacgcgcaccacttaa1680aacacacatttcaattggcccgcacaactgcaccatcatagatattaataaagcccaatt1740cgcctttgataactacacttctcaactccctaactcttgtgttcaccgtgccaacaaagc1800tgtttgcgccggtttgaatgctgatcttctttctacccactttaactacctctctaaagt1860agaaacatctaaaggaacttatcaatgggtacattttagccccaaaactacggcccctgc1920tatccaattctacgtcttgcaatttcagaaccacgtctacctacgtcttctccccatgcc1980tatgaacctttctttccgcactatctgcgacattgaaaccgcccacggtcaaccacaccc2040aacttacacaagcccagactggcgcgtgtattacctcgcaaagtacgatcccaccaaagt2100actcagagttcaagttgttgattgggcaggttcagataccattactgaagatcatgtcac2160cctcttcccacgccgacacattatgactaaagttttgaaaaacgaccaaactaaagttaa2220ctacatcgacccaactaaagatgcagcaacaacacccttcacaattgaaacagttgaagg2280aggcaaagttgaagtactctccgccaacgtcatccgccaaattaacattgaaactggtcg2340cggcaagaaacgtaaaacagctaaattagatgtaaaaatttcaattagagttaaagctac2400taaaacccgcacaacaattcacatcttcataacccaagcaaaagatgaaagcaaaatcgt2460cactccaaccttagaaatgcagggagtagtcgacgttttaatttccagagcatgtgtcct2520ctcacaacacgcaggacctgacatgacttatgaactgcacacacctgacggttcttgcac2580taaccccctcgttcaattctttgaagaagaaggcgaggttcctgaagatgtttacgatct2640cttcccatctgtagaacgtaacgctgaaggtttagttatgtgcatgccagaaggttttgc2700cctcatttctggttgcattcctcatccacttgcttactatgctgacaagtacggtcaaac2760actcgtacctgaattcaaagtgacaagaactcacgttgaacgcggccccattccaccatc2820acggtccatttactacacattcatcccaattaatgtagactccatcccacccttcggaca2880cacctacaaatccctcgccaaacttaacacatttaacctgggctgggaagatcaagacat2940gttagataacgctgacaccctccttccatgtctcttagaaatgaaaacatgaaacttcac3000ctaacatggaaaatcatcgcccgtgacttctcagataacaccactgaaacagcccttacc3060atctacactgattttacaactagtgatttcaccccagccaaccaaacaggatccagctct3120caatcccttagctctccacctctttcccaactctcttcccacaaatctcttgctaacctc3180tcagaaactggctcaatagataacatcccggaaaacacccctgaaacaatccaagaagaa3240gaagatatcgaagaggaagtaacatctcctgttcaacccacactcaacacactccaagaa3300caagctactctcaacgaatccattcaatcactcgttgaatcagttgattctgatgtccct3360tacacctgggaacaacgtgttgaagaaagtgaggttgcgactattgatttacaagctgta3420gaattagatcaaacacccactcccaccactctctcaacagaagcagtgcaagaatgggtc3480aacgcaacatcacctgaccttgctacagtagactggaacagtgcaataactgaacaggaa3540accacagtaacttcactaatagaagaaatctccctacgccccaacaccaacaacaccggc3600tataacttgtttaaacctaaccaacgtaacaacaacagcccatcagactacctatcacga3660acaacatcagcaaccaaacactgcttaattaacggtcttacttcaggttccaccatcctt3720gaagtcggatgcggatatggtaaagaattccacaaatacaagaactctaacatcaaagtc3780gttgctttggaacctcaccatgaacgctacaccgaagccgcaaagaaactaccaccaaat3840tctaccatttacaacaagaccctagaagaatacaaccaaattgtcattaaagaacaaatt3900gacgccctagtttcctttaacgccattcattacagtacagttactgaagctcttaaagca3960tgtaaccttgccaaagatcaagttgtcattttaacaccaaatcatgataaacttaaagct4020cttcacgatagtaacacaaccatcacaaattacaaaattgaacttgctaataacaaattt4080acattcacaatcggccaaaacacctacagcgaaaactacttctcacttcaggatttcatg4140attggaatgcatgccagcaacttcatgctttcttcccgctcttgtactctgcttgaacat4200gttaacacctttaatgcccacctcttaattgacgccacaacagaaacaaccaccctgcta4260gacgcctatgaagtactcgtcttcataaaatttagaccactgctcctcacctactccaac4320accgctcttgcaactaacaactcaagaacaatccgcagtgcactctcacgtcttattaac4380ttccgtgtcgcaccttctaaaacaatatcacaggtacatgttcacatcctcacccgcaac4440gaccacgcagccacagaacaagaacatctagacttagcaaatattgaaattgcccacatg4500aacccaaaaccagcaattatctttgtaacttcttcccgatccaaacataactacaccacc4560ccaactgtcttcttcgactttgctaaactatcagcccccaccctcccagaacagcttaaa4620ccactcactgatctagccctcgcaatccaacacactgctaccaaaactccaacaacagta4680cttacaactgccccagaggacccaaaaaccctcccaacagcacccacctcctttacaaaa4740ccttcccgtaaacgctaccactgcttccacgatttaaccaccctaccagttaacaaaaat4800ttcatcttagttaacgcaggtaacccccacttaaaatcaggtggtggaataactaaagcc4860gtctataatcgctacggccaaggattcatgcaaactaacaacagcatgcatgctacacaa4920ttacctcttgaagttggtgacgtcttggaatacccctctgaccgtaacaacctgattaac4980gtgacagccccctgcatcaacatcaaacacgcagattacaaaggcccacaaccagcccac5040gcactactagcaacatataacgcaattatcaactacgttgaacgtaagtacgataatggc5100gacgagaaacacgtcttcattcctctacttggtgctggatactacggttggtcacagatc5160caaagccttcaagcttttctatccgcgactgataatagtttccgcaacacatatgttcta5220ctcctacacccatcatctgtagatgttgaaacagctgaagtatttgcaaaaatgctcgcc5280actactaacccacactactaccatgttgagggcatagctaaagttaatgccaactcttgc5340gttctgcgtgcccacattgaccttgcccgcaaaacatcttaccctcaacctagcttagga5400ttaatcggcgcttactggatgtacacctgccccaccctggccgccccattgatacaagct5460ggcatgcacggtcccaacacctggactcctggcagctttattgacactagtattgctgaa5520atgactgctcgcgcttacaaaataaatcttgtttgcgacggttgcttcactgatatcatc5580aatttcaatcccgaccaaggtttaatctgcaatacctgcaagacacctgtcactctccgg5640acaaactattatcgcccacgcctcccacacatgattaaaaatagtcacggacaacccatc5700cctaacattattctggatggaatctaccacatggccacgcaccaattgttcctaaattcg5760gtactctttaaaaattcacatgctttcacaattaacaacctaagtgaatgtgatctccca5820actactatagcagtagcaacctacttttacaaaccttcagatgctttaaacttcttcctg5880aactttaactctgctatttctgctaacgtaacttatacatcagaacagataacacaagct5940caaaccacccctcaactcaccactgatgatgactactcggacttggacattgacacaatt6000aaccctacacctactactttaccacgcgccccaacaccgctttcaattagtgatagtagc6060gaagatatttcactctccgacgacacccacaccgtagcatcccatcacgcttcacaaatg6120tccataacagctgacacaactagtgatagatatagctctacaaactctctagctatctca6180gaagatataacatttcaaacaggtgacggtgaaacaacaacttcacttctgactgattcg6240gtcctatactactctgtcgttacagtcttacaactacgtaaagcagaggattgtgtagaa6300cctgctgtagaaactattaacctaagacccaactcatcacttcataccttgacacatgat6360acaaacgccaccgaccaactttacatccaagtggcttctgatgcttcaaatcacactgta6420atcaataacaccacacttgaatctatttctaaaacgatgtgtgtgaaaaactccttgtac6480accctacttaccaactcaaagtgtataagtcaccacatcatagttacaggatatgaccaa6540catgctcatgaaacacatactatcaaaagcgtcacgcgccaagccatgctcgccatccaa6600cccctttcacaacttacgcccacacacgtttacagcacacttcaagaagatatttccacc6660atcttccaaactgtcgtcctagaacacgtcaaccacaccaagctttccacttacaacacc6720aagcttcactacgcaaccacagtaccaacaaacctactcatcgatggaacaatcgaagat6780gttcaaggtaagctgcgcgaagttgatcttggcattgttgttccttaccagtgtgaagtt6840gacgagcaaattgaatacacaaatgcaattggcgacgttctctaccaaaacacaaatcac6900cactcaccactagtgacctttgtttctatcgacaaatcttttactaaacgtatttttaat6960tctttaaagtttcaaactctacagaacctatggctctaccttattaccctacaagtattt7020ttcacagttaaaggaattggcttcggcttcagctcaacggaaccaaacacacaactcatg7080aacggtctctggatacaatacggcatcttctacttccaagtacacctccaaaccctcatc7140acccacccagtgaaaaatggctacagactactttcattaccaatcatgctatacaacaaa7200ggcaaaagattcaccgaacgttcagtccaaacaatgacgggagaaatcgttaacacacga7260aaaccctcgttatcaaacgctgtacaaaaatacggcattaacactcgacttgaatacgtt7320gatatcttcgatctcctccactatacacctggatacatcgcgtttaacatcgcctttttc7380tatctctttgctggtaatggctatataacattcttagcctacatcatccaatttttcacc7440gctcgcgatatctacagactctctaaacacctctacacagtctcaaaactcaccaccttc7500ctcgcgaaatttaaaacagcaaccacccgtcacgtcttgtacgccaccaaccaagtaaac7560aaccgttcccttcatctcaccctcggatccacgacaattgacttactcagctttacaagc7620aaagcagctttccttgccctgtgctcctccttcatctcatggcatttcggattagctctc7680gccggtggtcactctggtgaagactactctcaaccaccctaccacgcagtatttcaacat7740gcactccgcactttcggacttattcctcctaaagaacattactacagctacccatcccta7800gcttcagcctgcgcagactcatcagcccctctctgcttcctcggaacaccatttaacttc7860cacttcccatcatcctacacacaagcagcatccatgactacagaacacgtagtaccattc7920tggatccaactaatcatcctagccccaccatcgctttttttagttgtcattccgtggctg7980atcacaatgttcatctcaccagtattaacatttaaccaacttttgctccctgcactcgcc8040gtacatttcttagctatctatgtaatctacctttactatttcaagaaatcgtgctgccat8100tctcacacatgccttaaacacgctaaattagaccgccccatccaaattaaaggcctcttc8160gatttatccatgtacttcaatcacaataaactttgcccccgccataacttttactgtaat8220aatgctgacgaacctggccacacacttggacccgtcctagcagctcacatcactgaattt8280tatcatcttaaacctggcactattaagcccgatgtcgccctagtacacacagaacaccac8340gcttccctgagtcgcaaagaattgaagaaacacgacgacccaaccgattgtgtctggacc8400gccccacaaggcgctgctgatactctcgtctacgaagcctactcccatttgaaaggagtc8460caaattcgcattgatgccaacaccgatacaacaactcgtagcatgcctattcccatgggc8520gccaaaacaacatcttctccagctacaggtgtcatcaacacaaaagttctcggccatctt8580aaaagacattgtaagaaaccagctctccaatcttacctctcaggatacgtcggtgccccc8640aacgctaaaactactatcatgcccattacgttttttaaacatctgcctgaaggactcaag8700aaatacgccatcaaaaacatcaccttctcaactaacatgctaactgacaactctaactca8760ggtttcatcccagaaggcatccacaacccaaacaacctcccaaccaagactttcctggtc8820aagtctatcgatcaaacaattttcgaccaactccacccagaaacacaagctatgctaaaa8880gacgaataccacattaaacttcatgatataacacctcaatttcacgaagttattgcttca8940gtgatgtcctacgaaagtttcatcttcgcactcatcattgtcttatccgtcttcctcggt9000tttatctcctacagctctactataactgccaccacatccctacatgccggcttgaacccc9060actatgcgcggatccaccacaggtaaaccattcattcaaccagttacttaccccaactcc9120attctaatccctctgacatcttcccaatataatgtattcagactacaaaacggaaccctc9180tacttcacttcccaaataactgccccaacctgcaccgttaaactaccaaacccctacatt9240ggcttaaccacagaaaaattctatgaatgctcatctgccctcctccacatgcccttcatg9300gttcatctcggtggtgtgaaaatcgcttaccttagagccggtgttgcttacaacatgcca9360acatactcagagacaccacaaactagcaaactctgtgttctcactgcactctcaaagacg9420aaatgctttaacatcctaccaggtggctttcatactccagtactagccatcctcgtctca9480acttgtgcacttctggcaacggtccttttctacctctacctactccagatctttaaattc9540tacactaactcagtcttgatggtggttgccattcaaatcgcaactatcttcctcttcatg9600ctatccccagctcttgctctaacacttcaactggtctgtctcatgtacctttaccataac9660tggatggtcgccatgattaacatcgcaaacctcgctgttctagcccacacaacatttggc9720atggtactcatcttcatctacgcgctctacagactctacaaagtctaccaagttacatca9780accgctgcagttacaattaacgcaactggtgaaatgcgcttccacggaactcttgatgaa9840gttgctctctgcacctttccaattagctctgcaagtgtttaccaaatcatgtctgacctc9900agcttagattacattaaactggccagttacaaaacatcttataaccgtgctacccgcaac9960ctggttaacatgatgcaaagatacattctcgacacctccatcccagctatactctatgac10020ggctcagtaacaactggcatcatgcaacaatccctagttagacttaaatccctagttaca10080acagtttacaaggataactactccaacatctgcaaaatttcatcaacaacccactcacga10140tcaacattctgcgtcggaaccttcctcaacccaaccactgttatcacttgctctcacgga10200atcttagcaaactctactatcaccgtttcccacaagtcaactatctatgaagcaacaatt10260gaatcacaacgtaatgacattacaatgcttaaacttacaacaccaaactccgcaatcact10320cacaccatcgagtgccactcaccaactgccactgacctcgaaagcatgcaagttaattac10380acccaatttgttactttcgctgataaagatcactcagatgccatagccatgaataacaca10440acctacctaccatctggtcacttctttgctatcggaaccgaatctggcgaatctggcagc10500ccctacttctttaacaacaaacttgtaggaattcactacggagttaacacctgtgatcaa10560tttatgctcgcctctaaacctgatggcagttactacatcccagaaatccaacacaacatc10620tccacacaaccatcttttagccttagttgcatagcccatggtcgaaaatcggtaaccgaa10680aaaaacacccccacacttcacgatgctatcaccaaagcccttgcccaaacaacacgctca10740gcttacactaacgatttaacaacagaacacacacctgaacaactccaggctgtccttgat10800tttctgacagaccctctgaataaatctggccctaagacactccataactacatggcaaca10860tccatccaagttcaatccaccaccacagcccaatccgcccacattgacgtccacaaacca10920attagacaagcagcaactctctactacctaatggttatcatctttgacatcatcgcattc10980ttcttaacccccggtgctgacttttccaaccaacttgtatccatgactgcctacctagct11040gcctggttccttaataaaacatgcttctacaacacccatgtcctctacacagtaaccaca11100tgcttccaactttgcaaatacttcaacattgtctacctcttcacaatcattcaatcctac11160aaaccaattgaaatgtcaatgatggataaacaatatgacttcctctgcgccatcattgta11220gccctcctactctctccaatcctggtcctcatctggaaacgcctctgcaacttcacctgg11280cattacctcgttgctcacctagttgttggctgtgtgctttgcatccacgtcttcttcagc11340tacagatccttcacatcttcaacaggcggtctcttaacattctcagaattcgcctttaac11400tcaccattaacagcagtcataacagctacagttgatgaaagtcctgtttctgttattgtc11460tacccactcttgcagcatgtcatcagcgcaaccctgatcttagcaatcgtgccatctccc11520ctcctcgcagcaacagtctactctttctatctcattgaccaagacgtcctctgcctccta11580atcttcctcctgcttacatccaaactgacaccacttttcctccgcccactcacaacattc11640gtttcctacgatgttgtcatcctctcaccctctgcttacgctcaatggctcatggcaaaa11700tccgaaggtaaagaatacggtctctttttcgctgtcattcaatcagtctttgactttaca11760tctcaacacgaagttgaattgacacttgaagcacaagagattacacagcaagtccgtgtt11820gaagttcataacaccgaacctaaagatactaaggatgacacctgctacatcacacccact11880actattattaacccaactctaattagaaccgacaattacttctacaccaacaccaagaaa11940gtaatctgcgaagctcccattgaactatcagataaggctaaagaaatcgtcgctaaacaa12000ctcgacgctattccacaccctgatgcagaaacagctgatctagaagttgccacacttatt12060gactgcacaactacaaccaagcaatcagaagttaaagaaattgaagtagacgatttaaca12120catgcctacctactcgctcttatcaaagcaacttgcatagaatctggcctccccattcca12180gtgatgaactacatctcatcccagacccaaattgactcagtgatgatccattactttgtc12240aaagctgccaacaaactactgcttaaagatcaggaactgcctcttatcactgcccaatct12300gacacctccaccattcaagaaaccttggaagctctcaaattaaaggacaccaccctaatg12360aacccccaagaacttaaagctcaccgcaaacagattaacattctgaaagctgaacaagaa12420cgacagctatccatggaacggaaactcaacgctttcctagacacaatgcaccaacgtgct12480gtggctgaaaagaatagatcagaagttcacagcaaagtgtcagctgctttacgcatgcac12540ctttcaaaggttgcaaacgctgcacattgtgacatggaaacaacagacggtgttatcaca12600ctatgctctgctttttctatccgctcagtctgcattactcaaaatactgcagatgtctct12660atctactactcacaacccacagatacatactcaatcacaactaccaccacagtcttcaca12720gcttataaccctatgaatgccaacggacaacccatcattccaaccactgacccttacaat12780cctatcatttcattcccggttatcttctcactaaaacaccgtgaggaaaaatcggagata12840gaacaacaagcaaacgtgggatactccgaacgtgccataaaaataacaacgactgaacac12900gccactggtacagttctcatgattgacaacattacatctgtcatccacgccactaaaacc12960gatgctcgcttcaccgtaaacggtaaaggctacaagatcgtcgacaatgccgttgcccac13020aaagctcgtgctaacatgtctacaattcttgctgtcttgcgcaaatccaccatcagcgca13080caagcagcagtagttagaattggcggtgctagataccaaacatctcaccaagctatctct13140ttcacccccttccaaacaccaggctacgttacctacgcaggaatatgcgtctgccaacgc13200tgcacactcagagaacaacacgagtgccactacgcaggtaactttgtccagatacctacc13260aatatctcaacacaaggacttaacaaaatctacgaattaacatctatcggcccatgtaaa13320cacaaccaatttgaatgcaccacatgccaaccagacttaattacaaaagcccccggacaa13380gtactggtatacttcacagcccacgcccaaacccactacaaacaaatagttactcaaatc13440tgcaaattctaccaagttaaacaatgcaactcccaaaaacatgcagactttactatccac13500ataattccacacggccactcaactgatacaactcatcctaacaccaccgttctagttgta13560cgaaaacacctagtcgatccagatcctgaacctccacacatcgggataagttccacctta13620actggcgaagaactgattactcttgacagattactacaacctgctgtcgagcctcaaact13680aagtcgtttttaaacttctggagcgggtggtaggtgtatcgttaccatccgctagcgacc13740cttatacatcagggagctataaaccgatacctcgtatcttttacattaacgttgcaacac13800gtaatgacacaaaattagataaagtctccaaactcaacacctggagcatggaaacaccaa13860gcggaattatcatggtaaaaaatcgtgacgcctctaaagaatttgatatagctaacaacc13920taccaaagtcactaccaattttaacacacaaactcatgacttacaaccgcgacaacctcc13980taatcagatactacacaacaccttactcactaggagaccttgtctacgcttaccaactcg14040gcgattttgaacaccttttactcgcggccggactttcaccaaatcctgcccaattgctcg14100accctggtaattacacccaatatcaacaaacaaaagaacatctcagacaaattatctaca14160gatctaatgaaaacctccataaaatcatttattacttcaccaccactgataaatatctgc14220cgctaaccctggacaacattgatcagaacggttatttgtatgatttcggtgactacccag14280ctaaaagtcccgtcaataatcaccacgtactagaatcatggacacaattctgtgtattct14340gcgaagttaaccttgctgaactaccaggttttaacctctacactcaaacccactcaactc14400cacaaattataaagagccgacaagacctagtctctggctcctacatggataaactactag14460ctcttaaccacctaatagcagcaaatccagaagccccttccaccgcttccactcccttct14520ttaacgcctccgcagccacaactgacatggtcggcctacactcaccactctttggctacc14580atgaagtcaacaatgatttgattcgctctaaactctcaccaacagaagttgacttcatca14640gatctcaagattctgcaaactatgctaaacccaacgtcatctccatcggcaacacaatta14700gctactacggcaaaagaataacaactaaagctgcccaaagaaaacctcaatacgatctcg14760acatgctcaaaagattagaatcaattggcatcagcacactcgctacaacatcttaccact14820accaagtagctgacgatgaacaatgtgtgcaagattttaaatactacgattacaactcgc14880ccaaaacctatgaccctaacttcctcaaggccatctaccactacatgcaagacatctttt14940ccatttacaaaactgatgtttcccttaaccatgaaacaggacaaccacgcccctcttcta15000tgggtcttggaatcacaggttttactcaaaaagctgtttggcagtctctaccacctggtt15060ttggcccacgcatgcttgaagttgcaaaagaaactctactaccatttacaacaaaggtaa15120ttgcaaaatacgctcgcacaaagaaagcccgtgcacgtacagtaggtgcttccagcttcg15180tcgcatcaacagttttcagatttcttcacaaacctgtaaccatgcgcatggtcgaacaag15240cccaaaacaacattggcccactcctaattggcgtctctaaatttaacggcggttttgata15300aatacatccgtactcaacatccaaacggtatagaacattacaacgtcatgggtgcagatt15360acacaaaatgtgatcgtagcttcccattgatcaaccgcgccttcgccgcagcccttttct15420atgatttaggaggtttaccatctgacaacatgtggtttatcaacgaaatatccgccttca15480tgactgacttctgcaacatctcaggtcacacatttaataaaccaggcggcacaacatcag15540gcgactcaacaactgccttttccaactccatctacaactacttcgtacacctcactgtac15600aattcctcaccttcctctcagccgacgtgccagaaacatacaaacatctccaaggttctg15660ctcatcgcgcttactcaactggaactctcgaagattacaacacctacttcgaactcatcc15720atgatttaaactgcactgaatacttctttcactttctcagtgacgattcttttatcttct15780caacccaagaagctttccccatcttcacaacatctaacttttcaaccaaacttcaatcta15840tccttggctgcactgttgacgtaactaaagcctgggctgcatctggacacatcgaagaat15900tctgcagttctcacattgaagaaatcgacggtgtctatcaatacattcctgatcctaaca15960acatgatcgcaggcttgtacatgtccggtgacccagtaccagtagataagaccatctgga16020aactagttgctacactcagtgaattggccatttttcaacacaaggacccatcacttttta16080acgccctttaccaactccttcaaaacatgcacagtgaatatgtcactcaacataaccacg16140ctctcctccctcttcaattgcttaacaagaaattctacactgatctacttcaaccaggat16200ttgacactgaaacaattttcctcactgacttgctaacatctgtcggtgtgaaactccaag16260catccaacgattcacaatgctacttctgtgacaaccctacaatttcaacatgtgctgatt16320gcgtagtgccataccccatgtgtgcactctgctcttacactcactgggttcaaacaaacc16380atgccccatcacaagtaccaccatgtgtgtcctgtggctgtgtcaatcctggaaaacttt16440accactgcttcactggcagcgttaaaatttgctgtcaagatcacgtctctggcctttccc16500tacccctagtagaccacgccaagaaagtaattaggatacccctcttccaaaactgcgaac16560gtcaacgatccttagtccatgcaataacacacacggccctatttgacgacaatgacgaac16620cacttaacactgcaacattttccaaatatgacccagaacaaactaaagaagaaaacttca16680tacgcatcctaaacgactgctacatgctcgaagcctaccaagcccaccaaaacccagcat16740acacctacgaagttactgaacctggcacaatagttgtatcacaacctccatcaacagcat16800atggtaactgtgtctacgctgaaattctcgatgacaaaaatcgcgttgtcttgaaagtca16860ctcttgatacaaagtcagtactaacacctactgtctaccatgtaacatcttgtcgtggta16920cactctacactcgctacaacaaaattcgcagagttcagcacaaaccttcaatagtcccta16980gatcaaccctagcctactttaacaacgcacaattcataattggaccaccaggaacaggaa17040aaactactttcgtcatcaagaactacattgacagagcagaaccaactaacaaaatagcct17100acatcgccccaacacacgcactcgtacaagctatggatcaagccatcttcgaagctcacg17160gatatgcagctagtgtcaacattgttaaatcagaacttaacaacaagacctacaactaca17220aagcaaacatgcctggatgcgatatcatgctcggaacacccggcgctgtatcagcacaca17280ctggttgcaccatgatctttgacgagatgtcacttgtacagttgaatactgctatgactg17340caattgctcgagtaaaaccatctaaactcatcctgctcggtgacccatttcaattaggcc17400cagtagttcactttagaaacttccaatacaactacgaagacttcccctatcttcatttcg17460ccccaacaataacatctctccgcgtatgcttccgatgcccaacccaaatctttaacgtat17520ggagtcactactacaccaaacatgacatcccactcaatgcctttaaagccggcggctcgt17580ttgaaatcattaaacaccactcatgcgccagagtttcagaaacagctcaccctctggtca17640caagactctcagaagaaaatccagactccatgatcctttgtaactacaaagctccagtct17700cctgcctcgaaaacgctctaacaattgactcaagtcaaggcagaacctaccgagacgtca17760tagtcgtgttgctcggaaacacaaatttcacccgtgtacccaaccgatttattgtcgctg17820cttcaagagctacacattctctcaaagtttacctttctgaagagatgtacgaccactact17880ccacactattcaattggccacaattagaaacaccacaaatccagccacaaaacgccttca17940caacacctaacaacttagaagaaacaaatctcacaatggccgctaataacatcagctccc18000ttgtcatctgtgatatagaatttttccatcaagttcgcagccttgaacccaaagtgtgca18060cacttgaagtcggtgacattaccatccacacatcaacaagcgctaaacagattatcgttc18120cacgcgtttcatcaacatctagtgaaatcttcccgaaatacttcgctcacaacatgccaa18180ccctcactctccccaaaccaacaccaaagggagattgggattacatgaaacctcaccgtg18240gaatttctcgccgcattaacatcgacagaactcacaaactcttctcccatatggcttgca18300caacaaatggaactcccatctatgttctttacggtgcccgcaacgatatcctagcccttt18360ccaaattaaatatcgttggtgactataaatgtttctgcaatctaccagctaactacttca18420ccctagatgcccaagataacatcaaatactactgcacccaacacgccaaaccagcagcac18480ctctcgctggaaccattaatgctaaatacattgacatccaaacagaaaagatcactcttg18540ccaaacgccatgctgaaatttgtgatcaccaccatggtgcagcacatacatctgatgtcg18600acaccatcatgaccggatgcatcttaggcaatgaattgacactagcaactcaacatctca18660gccaacttctcttcacggaacccttcagccactactcaagtgtaatttctccagatatgc18720accttacagtttcaacccgcccttgcaggacactcggacaacttatcataactaacatca18780atggtaagaactacagattttcaatccaaccacacctggaccttaaaacacactttttca18840acatcgttgctaaacaaggccactcctctgttttagtcaacgttccagaaacagttccga18900cctgtactaaaattcgcggtgaaacctgcgctacatgctacaacacctggcttatcctcc18960aagaacttcacgctgaactagctaacacaggtctcgccctggttaacccactccaacgcc19020aatacactcaacgagaaacacaaattttaaccaaggtaatgtctgtcactccaacatcta19080caggcaaaatcatggtccatctcgacaacacactctccttccccttcctaatcgatttac19140caacaagtatccaaacatactgcaacaaaattgataaaccaatccctaaccctgcaattt19200tcacttccctaaaaattacacacacctacggatttggaacaccatttgtccctatgacaa19260acaacccactcattccatctatcctcagtaaccaaaaacttgctgacaacccggactact19320actacatcacacatcaaccatcaaacagcgaactttctttcatgactttttcgccaactg19380ctggcttctactgctacgacacccctctgatcaacttctcggaacgtaccatcaataaac19440caccctactatatcacacgaattgtccgtggccaactacaaaaacctgatattggcctct19500tctccactaatagacttttcaccaaccaatacgtacctttacctaatgaacaacaagcgt19560acgaagactttggtaaacatgtctcacttggcgaaacaacatcaacatcctggacaatcg19620gtggcatgcatctactcccagccttccaaaccacaaccaattggcagatcacatcagaac19680atacaccaattctctcaattaccatagctcgcgatcgcggggtcaaactaggaacaacag19740ccttagatgtacctgctgaccaactacttgaattgattgactataaaaacatcacagtct19800caaaagtaatagctattaagatcgacggccaaaaagtaaacatgatggttttcactcaaa19860acggtaaaatccaaacagcctacccagtacaacaagcaaagattcacgatgtgacaaaac19920accacgtcactcctgactacataatctggcctaaaattttcagcgactcacctgtcgaaa19980aatgggatcttcctaattacaaccaaccacctaaaggtctcacatcaaacgtcaacatcc20040ataaatacgatcaaatgtgtgcattctttgacaaacacctaaccattccaacaaatggcc20100acgtacaccatcttggtcatgccggacaagtttacagccctggtgatatagttttaaaaa20160gctacttcacaaaagctcgtttaacatcatacgacttgcgtgattgcatatcaaccagcg20220aaacagctcaccctgatgacaactggcgtgcacacttcatcctgtcagatgcatactgtc20280ctaacaccgattttacatctctctgcaaagactacatgactaaacacctacgtcttggcg20340gaagcatgctctggaaaatgacagaaacctgcaacatcgacgctgaatccattctaccat20400acttcggatctttcacctttgttactttcgccgttaactactcatcctctgaaacttacg20460ctttccttaaggggtacatgggtgaacctaattacaatactacactcccagctaacatca20520tgggtcaactcggcgcgtacagacaaaacctattgtacgtcccctatgctagtgagtata20580caggtaccagagtttataaagattctggtgctgtgaaagttattccggccatgtatgcag20640atcgtctgaatacttcccactatgctctcggctgcatctacagcacttagacatggcaaa20700cttgctatggctgactcttctcatcactctcagtctcgtgtccgctcaaattaacacgac20760acttccagactgtcctaccccaacctgcacctacacaccaccaacaacgaccccatttga20820ccacacatcactacactcttttttcacacacgacacaaactatgctttccttcttgctct20880taagtataatggtactttcattcctgccactatggctatgcctgctaacaacctttcgcg20940aacatatatactcaactataaaaatgtctttgtcaacgtacctgattctcaactcccccc21000tcctcaaagaagtaatactcagttggctgttgttgcaactacagcttcctgtctcactgc21060aacgtatactgcgattcctggtactattcctggcctttcaaccaatcaaatgtattgtgt21120gaaatttactcagtcgtccaactttgtgtacttgtacgataccactgtgaacacccctgt21180tggcgtacttttaaaaaacccaactgccccagtagttacaactatcctatccactatcat21240ggttaaccgccctacaacagtttcaacctcacagctgctagctaatatctactcccttat21300gagaacgagaagcaacgtgattcaacaacaattcacttgcacatcttgcctctttgccat21360cgacgcatctacaggaagagcagctacctcaccaactacttacctcctaaacaactggtc21420cccatccttctttgaaactaaaacagctatgttggaccacgtctctatgaacaccatcta21480ccaaacagcacgtggtgttactcgtgaaaccttcactggttacatccttaacgcagctaa21540aaccgctgcctacgctcgtggttctcccgcctattacatcttttcattcgcagccattgc21600acgctaccatcaagcaatggttactccttataactataacatcggaaaactttgtgaagc21660aacagctgcttgctgcttatcaaatggcattgaacattttgacatctatacctggctatt21720tcactactacggctttatgaaatccaaagtctacagtgaaacttttacaacaaacccagt21780actaacatcagttattaacttagccacatcaattaaaggtaaacattctgtgtactgtga21840cctaacaccttacagttcatttactgcctataatttgatcatgcttcaagatgttgtcga21900acaataccaagggacacgtacaacatccacatacgctaacaattttctcaacctcactga21960tattaacggatacctaccctattccaccgatacccccctcccacaaaactaccaaaacta22020catgccttttgcaacctacctacttgcaacattttactcttctttcactttacaacagaa22080atttgactttcacagattcctccaatctgtctgttattatgactctggtttcatttctcg22140taaccaagcctaccattcgatctgtcacataatcactctgaaaacatcatcttcagtgcc22200ttaccccgcagaactccaatggccctttactgctctcccttcctctggttatgactctta22260tccagtttctacagatttaggcacagtagctgttggcaacacaaatccacttgattaccc22320tggaatttgttaccaatcagtttgcttgactcatcctactattaacttcagacttttgtt22380atcagaattaccaattaaatgggaagtccgacctgaaggtatctatgtaacatctagaaa22440cgatcctacatgcctgaatttcaaaacatcacagttgggaacagttttcattaccctcta22500cgattctaccgctacatgcacactccaacagactacttactctcgctctgctgtcctgcg22560ccaagtcaacacgtttacaccaaatgctactgatttagtcgtgccttacggcaaaacacc22620aatatacaatggagcctataatattaattccactgcatctggtgctgtctctctctctac22680caacacacactacattccacgtctcctcacttcagttttaacaccgcaaagtgactacaa22740gattacacctacaaccactaatcttgaaccattcgtggaaattaatggcacaacctttta22800ctacgatgctaatcaagcctacgtcaacacatcggacgtttatcaagctaacgttgattt22860tggactcccccaaacattttctcttgataccagctttcttaattgcacagctttcctctg22920cgcttctgactacaaatgtatcgccgacttttcaccaatctgctcagctacacaaccact22980aatcgacgccctactccaaacctacacactgtacaatcaacaacttacactgtacaattc23040ttccctgctcacaattcaagccgctgcaaactacctctacgacacaagaacacgtactag23100acgatcactcctatctactgttaaccttggctacggtgacgtttttcaacaaccagcctg23160ggtctacgaaataggtgcaacagctgcaattcctatgatcggccctgcaatctcagtcgg23220tttgttagccagtagagtagcatctattgaatcaaccctctaccaaattacccaaggagt23280gaaggagtctattatggctatgaacaccaagcttgataagattgccaacattttacactc23340agacattgaaagattaggattaacagttaacacattagcctcttcattcaacacattcac23400taaacaagtaacagataaatttgcctctgttgatgttgcatttaacgacttatcaacatc23460actgaacacactcaccacaacaacaactgttggtgcctcttatgcagctcaaattaactt23520cgcaacaaccacgctaaccaatttagaagttcaaatctctcaaacaacagatcacgctct23580atcttgcatcgctgcgcttaatgaacatgttctttctccacactgcatttccgtttcaca23640acttagcgcttacacacctaatgctctctctaacatcggtgaaaaactagtactaactca23700ctacatcaacggatcactcatcacattctaccatctcattgctagcaagggagtttacag23760acccctgcccaaaatgtataactctacacaccatttagttgcaacaactggcttttataa23820cggccattgtatcttctgtggtgaacattattgtgatatcagcgtttctgtaccatgtga23880ttatgcgtacgaaccacttaaatccactgtactagcagcttacccactaacaactggtgt23940agcccttcttactcttgactctacagctactaacttcgttattaactataataatgcccc24000tctcatttcatcaagcgttccacaaccacctatcatcaataacaccgatggtacagttat24060tacgctccaacaacaactaatccagctccaaactcaattcgatcactttaacatctcgtc24120gaattttcacgtttcagaaatccaaccagtcatagactactacaatgcacaaatcaactc24180atctctaaaccagatagttgatttcggaaatggctcaacatctaatgtaggaacaatcat24240cctcattgtaactatagttgcctttgctcttgcatcagctattgccattatctgtggatt24300aaagcactaaagaccaacaagttgcaatggtttccaccggtacaaacaccaccgcaggag24360atcccaaccttgggtttcacttacaacttaatgatgtccttaacaccctggacatctccg24420gcttcagcatctcaaattctcatttgatgagtgcgatgctgaccctcaaccttctctgga24480gcgctatctttctagtctccttttatctaatcttggacatggttatgtactacgttcccc24540tgctcaataagattccactgctgccaacaatcaaaaacctttgttggtttatctcgactc24600tgcttgttgctgtttccatttacgcttcaggaactctgctgatcatcaaaatcttcggag24660gcgtagtcttgctccttgcttttctcggcatggtgaagactttctatatgctcattgcaa24720tggcgctgtcctactaccgacaccgtaacatcgcagtggcattgaacggccctaatgtcg24780ttatggttaatggcacagcagtccactctgatttcatccccacgtctgtgacactggtga24840agaaagatggtgaactctattttcaaggaacaaaggtccccggaaacaacttgattgatg24900caaccgtgaagatcttcactttcaacgaatcgtttgaatgcctccccatcgcactctcac24960gttacactaaaggttcctacgctagatttgagctcaagtcgaccaacacccactatgaac25020tccaaactttctatccttcgcaaacagaaactaaatctgtttgaaacactagcgaaaaca25080aattacaatggcattctacggacaaccaatgatgcctatgcctatgcctgcttcctttcc25140tcctcgtcgccgcgcccctaaaaagcgcccgccacagaagaaacaagccaagccccaacc25200taaacctcttaatgaccaacagaaaacagctgttgcaaaagaggctattgccaaaaccaa25260tactgtgccaatgccacccatggccacagtagtgaagacaaacctgcgcgttcaggttgt25320aaaaagcaaacgcggtgacctcctagatgctcgagggcgtcttacacaatcagagatcaa25380ccagctgtatttccaaatcgccactgccctcaacaacggcctcggagaaattggaataca25440caatgcttcaacacaggacggcgttcgtctcatcgtgcgcctcaatttagcccctcctac25500ccctattgctgagaaactctacaaatctcacgcggcggctaatctcaactcagaaattct25560ggtttctaaacaaaaggaactggagcgagaccccatttttcatgtttcctcgactgagta25620aactgacttcacctcaaaactgaagtatatataatcacatctatcacactatcctttata25680aactatatattggggtggcattaggccattccaaagaattctttaagatcacactcctcg25740atacctgactagtcatcacgtttcctggggtctgctctttaacattctactatcctataa25800gatctaaaactatccaaaaaaaaaaaaaaaaaa25833<210>2<211>18<212>dna<213>人工序列(artificialsequence)<400>2tgcccaccaagacgacta18<210>3<211>18<212>dna<213>人工序列(artificialsequence)<400>3tttgcgctagcgcgattc18<210>4<211>43<212>dna<213>人工序列(artificialsequence)<400>4cgttccaaaccgtcttgggtcaagagtgacgtaactgctactg43<210>5<211>44<212>dna<213>人工序列(artificialsequence)<400>5actcacttctgctccttgcgacgcagagtgtaagaggttttcca44当前第1页12