本发明涉及一种基因突变检测试剂盒,具体而言,涉及一种易栓相关基因突变检测试剂盒。
背景技术:
近年来,随着中国人群易栓症发病率的升高,人们对其危险性的认识和重视程度也逐渐增加。易栓症(thrombophilia)是一种容易发生血栓栓塞的病理状态,一般由抗凝蛋白、凝血因子、纤溶酶原等遗传性或获得性缺陷,或其他获得性危险因素引起。患者的突出表现为容易发生血栓,虽动静脉均可被累及,但静脉血栓(venousthromboembolism,vte)更为多见,其中深静脉血栓栓塞(deepvenousthrombosis,dvt)和肺栓塞是最常见的血栓类型。vte的发生严重影响患者的生活质量,重至危及生命。虽然获得性及遗传性危险因素均会导致vte的发生,但遗传性危险因素携带终身,对其明确诊断具有重要的临床意义。目前较为熟知的遗传性危险因素包括:抗凝蛋白的缺陷,如蛋白c(proteinc,pc)、蛋白s(proteins,ps)、抗凝血酶(antithrombin,at)质或量的缺陷;凝血因子的单核苷酸多态性(singlenucleotidepolymorphism,snp),如凝血因子vleiden突变、凝血因子iig20210a突变。此外,近年来也有新的遗传性危险因素在人群中发现,如凝血因子ixpadua突变及凝血因子iiyukuhashi突变等功能获得性突变的报道。下面简单概述已发现的易栓症相关遗传性因素。
1.蛋白c抗凝系统缺陷机体抗凝系统的存在避免了凝血过度造成的病理性血栓形成。蛋白c抗凝系统是参与抗凝过程的重要组成。pc是整个系统的核心成员,它主要通过灭活活化的凝血因子v和凝血因子viii发挥抗凝作用。pc的活化需要血栓调节蛋白(thrombomodulin,tm)的参与,ps作为辅因子加强pc对底物的灭活作用,而epcr促进pc与内皮细胞的结合。因此,该系统相关蛋白的基因突变导致的蛋白缺陷均可能会造成抗凝功能受损,导致血栓形成。
2.抗凝血酶缺陷。抗凝血酶(antithrombin,at)是血浆中最重要的抗凝物质,具有广泛的抗凝作用。它可灭活凝血级联反应中的多种活化凝血因子,主要通过灭活凝血酶和活化的凝血因子x(activatedfactorx,fxa)发挥抗凝作用。此外,肝素辅因子ii(heparincofactorii,hc-ii)也具有类似at的抗凝作用,但其对凝血酶的抑制作用明显强于fxa。因此at及hc-ii的基因发生突变均会导致抗凝功能受损,促使血栓形成。
3.凝血因子异常。临床上凝血因子的缺乏一般会引起出血,但研究发现,凝血因子某些突变蛋白促凝活性增强,从而导致易栓倾向。据研究报道,凝血酶原f2基因3’utr的g20210a突变可增高血浆凝血酶原的水平,从而导致易栓倾向。此外,已有文献报道f8、f9、f11和f12等基因突变使得凝血因子viii、ix、xi和xii异常增高,破坏正常的止血平衡,易造成易栓倾向。
4.纤溶蛋白缺陷。纤溶系统主要涉及两个蛋白,即纤溶酶原(plg)和组织型纤溶酶原活化物(plat)。纤溶酶原在组织型纤溶酶原活化物的作用下活化为纤溶酶,这两者的缺陷引起机体纤溶水平下降,导致易栓倾向。
5.富组氨酸糖蛋白(hrg)缺陷。富组氨酸糖蛋白(hrg)为一种多结构域血浆糖蛋白,可与多种配体结合而发挥作用。hrg的配体包括锌离子、肝素和硫酸肝素、纤溶酶原、纤溶酶、纤维蛋白原等。已有研究证明,hrg能够抑制纤溶酶对血凝块的纤溶作用。hrg的抗纤溶作用会导致易栓倾向,但目前临床上无法开展此蛋白的生化检测,因此hrg的基因检测非常有必要。
6.adamts13缺陷。血管性血友病裂解蛋白酶(adamts13)是一种血浆金属蛋白酶,主要将血液中血管性血友病因子(vwf)多聚体降解为黏附活性低的小分子片段,它的缺陷可导致血栓性血小板减少性紫癜。vwf是参与止血的重要蛋白,其多聚体的大小与其功能密切相关,它的分子量越大,与胶原和血小板的结合能力就越强。因此,adamts13可以通过对vwf多聚体大小的调控来调节vwf多聚体与内皮下胶原和血小板的粘附能力,从而影响血栓的形成。
7.组织因子途径抑制物(tfpi)血管受损后,活化的fⅶ(fⅶa)立即与暴露的组织因子(tissuefactor,tf)结合形成fⅶa/tf,启动外源凝血途径。血浆中tfpi首先与fxa结合,直接抑制它的催化活性;然后在ca2+存在下,tfpi通过k1与fⅶa/tf结合,形成fⅹa/tfpi/tf/fⅶa四聚体,抑制tff/ⅶa对fⅹ和fⅸ的催化活性,发挥它的抗凝作用。有研究发现tfpi突变后会造成易栓倾向。
目前针对引起易栓倾向的相关基因突变,多采用聚合酶链式反应(pcr)扩增目的基因的外显子及启动子区序列,然后应用sanger测序的方法进行检测从而获得突变位点。该方法是参考genbank目标基因序列,设计引物分别扩增基因的启动子区和编码区,采用sanger测序的方法进行测序。由于易栓的遗传异质性和多个基因突变均可导致疾病发生,传统的sanger测序技术已经远远不能满足医学研究和临床应用的需求。而二代测序的方法应运而生,具有通量高、速度快、价格低等优势,并且可以同时分析大量样本而得到广泛应用。多重pcr富集二代测序技术是针对目的区域设计的多重pcr扩增体系,从而达到富集目的片段的目的,对研究者感兴趣的基因组区域进行捕获富集。
本项试剂盒发明采用了多重pcr富集二代测序技术开发由于遗传因素导致易栓症的基因突变检测,全面检测18个基因及3个基因特定多态位点突变导致的易栓症,临床应用前景好,特此申请发明专利。
技术实现要素:
本发明的目的在于克服现有技术存在的以上问题,提供一种易栓相关基因突变检测试剂盒,本试剂盒采用多重pcr富集二代测序技术全面系统的检测proc、pros1、thbd、procr、f5、at3、hcf2、f2、f7、f8、f9、f10、f11、f12、plg、adamtsl3、hrg及tfpi18个基因的全部外显子及调控区的点突变,还针对plat、fgg和habp2三个基因特定的多态性位点进行检测。本试剂盒旨在应用于生物学研究和分子诊断由上述基因缺陷导致的易栓症。
为实现上述技术目的,达到上述技术效果,本发明通过以下技术方案实现:
一种易栓相关基因突变检测试剂盒,包括:
ddh2o;
10×buffer(takara);
2×gcbufferi;
dntp;
mgcl2;
hstaq;
多重pcr引物混合液;
illuminai5/i7引物一系列试剂。
进一步的,所述多重pcr引物混合液序列如序列表seqidno.:1至seqidno.:774所示。
一种对基因组目标区域点突变的检测技术,针对易栓相关基因(见表-1)的突变检测体系设计方案如下(参见图1所示):
1、针对proc、pros1、thbd、procr、f5、at3、hcf2、f2、f7、f8、f9、f10、f11、f12、plg、adamtsl3、hrg及tfpi18个基因转录起始点上游500bp,5’utr,编码区,剪切点8bp内含子序列,转录终止子及plat、fgg和habp2三个基因特定的多态性位点设计引物富集捕获(引物序列信息见附表-1)。
2、将扩增出特异目标序列,加上index序列和illumina测序接头与片段连接;
3、模板分子与芯片测序通道上的引物结合,通过桥式pcr生成簇,以供测序反应。将测得序列与参考序列比对,从而找出突变位点,并对突变位点进行功能注释。
4、对所有明确与疾病发生相关的致病突变进行sanger测序验证。
表-1
本发明的有益效果是:
1、检测范围广:全面检测18个基因及3个基因特定多态位点突变导致的易栓症,较全面的检测由于遗传因素缺陷导致的易栓症,能够更好地应用于临床。
2、通量高:多重pcr富集二代测序技术可实现同时筛查数百个患者标本的易栓相关基因的点突变检测。
3、检测方法快速、准确:多重pcr富集二代测序技术,无需复杂的建库过程,而且利用illuminamiseq测序仪的2×250bp测序模式对目的片段实现双向测序验证,大大提高二代测序的数据准确度。
4、检测准确性高:二代测序所有目标区段平均有效测序深度>300x,所有片段测序深度>=20x,且对所有致病性突变进行sanger测序验证,保证数据准确性可达99%以上。
5、成本低廉:相较于传统的sanger测序,针对目标区域采用多重pcr富集二代测序方法的成本将大幅降低。
上述说明仅是本发明技术方案的概述,为了能够更清楚了解本发明的技术手段,并可依照说明书的内容予以实施,以下以本发明的较佳实施例并配合附图详细说明如后。本发明的具体实施方式由以下实施例及其附图详细给出。
附图说明
此处所说明的附图用来提供对本发明的进一步理解,构成本申请的一部分,本发明的示意性实施例及其说明用于解释本发明,并不构成对本发明的不当限定。在附图中:
图1为本发明突变检测体系流程;
图2为snv在基因区域上的分布统计;
图3为snv功能分类;
图4为部分样本sanger测序验证结果图;
图5为实验检测12个样本平均测序深度(横坐标为样本,纵坐标为平均测序深度);
图6为碱基覆盖深度比例(横坐标为样本,纵坐标为覆盖比例)。
具体实施方式
下面将参考附图并结合实施例,来详细说明本发明。
实施例:
利用本发明所述基因点突变检测方法对12个易栓症患者样本进行检测。
一、检测样本
将待测样本浓度标化至20ng/μl,样本体积要求20μl。
二、检测方法
目标区域富集测序方法检测点突变主要包括以下步骤:
本实验所需的试剂包括以下:ddh2o、10×buffer(takara)、2×gcbufferi、dntp、mgcl2、hstaq、多重pcr引物混液、illuminai5/i7引物一系列试剂。
a.反应体系配置
a)10×多重pcr反应体系配制:
10μl体系:
pcr反应条件设置如下:95℃2min;11个循环扩增(94℃20s,63℃-0.5℃每循环40s和72℃90s);24个循环扩增(95℃20s,65℃30s和72℃60s),在72℃延伸2min,4℃保存。
b)gc多重pcr反应体系配制:
pcr反应条件设置如下:95℃2min;35个循环扩增(95℃20s,68℃60s和72℃2min),4℃保存。
b.将pcr产物按照比例混合并将得到的混合产物按体积比样本:磁珠=1:1使用pcrcleanbeads进行纯化。
c.i5/i7indexprimer对纯化产物进行加indexpcr扩增
10×反应体系配制:
10μl体系:
pcr反应条件设置如下:95℃2min;12个循环扩增(95℃20s,60℃30s和72℃60s),在72℃延伸2min,4℃保存。
d.将pcr产物按照体积比样本:磁珠=1:1使用pcrcleanbeads进行纯化。
e.按照2100bioanalyzer标准操作流程对pcr产物进行片段分析,确定产物扩增片段大小。
f.样本文库进行上机测序前定量,使用qubitspectrophotometer对文库进行精确定量,换算浓度(nm)。
g.将定量好的文库混合上机测序,最终文库上样量11pm,按照miseqbenchtopsequencer标准操作流程进行测序,双向测序,两端各产生250bp序列,及两端各8bp的index标签序列。
h.通过生物信息的分析对各突变进行功能注释。
通过生物信息的数据分析可得到目标区域的所有snvs。
1)snv基因区域分布统计
这些snvs位点相对于基因的位置分类如下表所示,包括外显子突变(exonic)、剪切位点突变(splicing)、non-codingrna、基因非翻译区域5’端突变(5’utr)、基因非翻译区域3’端突变(3’utr)、内含子突变(intronic)、基因上游突变(upstream)、基因下游突变(downstream)、基因间区突变(intergenic)等。参照图2所示。
表-2snv基因区域分布统计
2)snv功能类型分类
相对于其他的突变类型,突变发生在外显子区域有可能直接导致最终产物---蛋白质发生序列以及结构变化,而这种变化最有可能是导致性状改变的分子机理。根据不同的变异类型,可以分为frameshiftdeletion、frameshiftinsertion、nonframeshiftdeletion、nonframeshiftinsertion、nonsynonymoussnv、stopgainsnv、synonymoussnv。
表-3snv功能类型分类统计表
3)snv功能注释
snv的详细注释可以使研究变得更加容易,通过这些注释,包括保守性分析,1000genome、esp6500、hgmd数据库注释,go和keggpathway分析,通常可以迅速的将相关snv的范围缩小到几十、甚至几个,在保证研究准确性的前提下,极大的提高了研究效率(部分展示见表-4)。
表-4snv注释统计
三、数据分析表
根据上述突变snv功能注释,12个易栓症患者突变检测如下表-5所示,95%的患者找到疾病相关突变,且经sanger测序验证均为真实存在(图-3为部分测序图展示)。所有样本所有目标区段平均有效测序深度>300x,所有片段测序深度>=20x,均达到测序质量要求(图-4、5)。
表-512个样本突变检测结果信息
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
sequencelisting
<110>上海交通大学医学院附属瑞金医院
天昊生物医药科技(苏州)有限公司
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tgcccacaattatccccctg20
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<212>dna
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cacctcctccagaccttgatca22
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gggtcatcatagggcacataatcaat26
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cactttctccagaccttggtgaga24
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<211>28
<212>dna
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ccaaggtctggataaggaaaagactcat28
<210>39
<211>21
<212>dna
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ccccagccatacaaccctttc21
<210>40
<211>22
<212>dna
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gggtggtgtcactgatgtctgg22
<210>41
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<212>dna
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gaactcagccatatgactctctctcc26
<210>42
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<212>dna
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ctgcaaagaggggcatctca20
<210>43
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gccaggtgaccctctctcca20
<210>44
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<212>dna
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ggtttgtctggctgaggtctaaag24
<210>45
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cctcttctccagagctcagtgaaa24
<210>46
<211>27
<212>dna
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tctgaatgagttctggagagagagtcg27
<210>47
<211>22
<212>dna
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gccctctatggattttggctgg22
<210>48
<211>22
<212>dna
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gggaaggacttgtgacttcggt22
<210>49
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<212>dna
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aaaagagaagcacacacaccatgc24
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gacctggaggacagcttgcc20
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tggggagaaagcacccctct20
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agagggtgaaaggtcctcggag22
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cttccagaatgaggccctgg20
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ggtggccactctgctttcca20
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<212>dna
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tcctctacagccagatgtcacagg24
<210>56
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<212>dna
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tttaagagtaacagatcactaggagggtcc30
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<212>dna
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gttggttcaaattattcttccccaagtaat30
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gaattctccagcaccaagtgaaagta26
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<212>dna
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gctgactatgattaccagaacagactgg28
<210>60
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<212>dna
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aggttctgcaaggttattgacagtga26
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<212>dna
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gacttccagatctctcttctgtctgtctct30
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<212>dna
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tcaatgatgagtttcggaatgacc24
<210>63
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<212>dna
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cataactactcttggattctgctttgatga30
<210>64
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<212>dna
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caaagcaagcttcctctgtgagtg24
<210>65
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<212>dna
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ctctcatttgaaaattctgagtttgatctt30
<210>66
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tccccacactacagaagtgcca22
<210>67
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gcacagtcatatttcaaccacagga25
<210>68
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<212>dna
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gccttatttctgagttctctattctgttcc30
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<212>dna
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catcgcctctgggctaatagga22
<210>70
<211>20
<212>dna
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tgccttcggcagtgatggta20
<210>71
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<212>dna
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ttggttccagcgaaagcttattt23
<210>72
<211>20
<212>dna
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gcctgtccagggatctgctc20
<210>73
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<212>dna
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cacatgtatacacacacaccatccaaaa28
<210>74
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<212>dna
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aaatgtggcagcctctcagaagtt24
<210>75
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<212>dna
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ggtttccccatcctaccgagat22
<210>76
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<212>dna
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<400>76
atgtaatttctcccatgattctgtatttgt30
<210>77
<211>30
<212>dna
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tcttgagttatttcattgtctttctgtcct30
<210>78
<211>22
<212>dna
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tgtcttgaacctttgcccagtg22
<210>79
<211>22
<212>dna
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<400>79
ttttggcctgtgtctctccctc22
<210>80
<211>26
<212>dna
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tctgcattgagaagcaagactgtcag26
<210>81
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<212>dna
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gcagtgctactgaaaacatgtctttcct28
<210>82
<211>30
<212>dna
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tgattttactgttgtttaatggtacacagc30
<210>83
<211>26
<212>dna
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tactcctccaggtgcttcttaccttg26
<210>84
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atgacagaactcctgaccattcca24
<210>85
<211>28
<212>dna
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aagatcaccatgttatcaaagcctttgt28
<210>86
<211>18
<212>dna
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gagccacagcgtcgtcca18
<210>87
<211>26
<212>dna
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<400>87
tgtttttcttgcttagcaactctgcc26
<210>88
<211>30
<212>dna
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<400>88
tgctggtattaaagacttagacattttccc30
<210>89
<211>22
<212>dna
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<400>89
gcatgtgaatgccaaattaccc22
<210>90
<211>32
<212>dna
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tgaatatgcctttcaaattagtttttgtattt32
<210>91
<211>18
<212>dna
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cacgcctctgggtcctgg18
<210>92
<211>28
<212>dna
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<400>92
ttgcaaagggaatgttctctaaaagaaa28
<210>93
<211>19
<212>dna
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cctcccctgcagttcttgc19
<210>94
<211>23
<212>dna
<213>artificialsequence
<400>94
aaggttctccaagtgtgagcttg23
<210>95
<211>22
<212>dna
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<400>95
agaaaaagcggagggagtgaga22
<210>96
<211>18
<212>dna
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cccccagcctacccagct18
<210>97
<211>21
<212>dna
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tttcagccaagctcttcaagc21
<210>98
<211>21
<212>dna
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tcgtccgttaccactgacctg21
<210>99
<211>22
<212>dna
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accccaacagggtgactttcaa22
<210>100
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<212>dna
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tgcagagtccatttataatgtgagatgg28
<210>101
<211>28
<212>dna
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atccaacctgacccaaatgtacttttta28
<210>102
<211>24
<212>dna
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<400>102
gttggctactctgcccatgaagat24
<210>103
<211>26
<212>dna
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<400>103
tcttcttatgtttcaaaaagccccaa26
<210>104
<211>22
<212>dna
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<400>104
cctgctgttcatgcatctcctt22
<210>105
<211>24
<212>dna
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<400>105
tgatgacatcaccatggtcctcat24
<210>106
<211>18
<212>dna
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<400>106
tgcgggtggagaagggag18
<210>107
<211>30
<212>dna
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cctatgaatgtttgtgttcttactttgtga30
<210>108
<211>22
<212>dna
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<400>108
cctctggggtgagttccttctc22
<210>109
<211>30
<212>dna
<213>artificialsequence
<400>109
agcacaggtgagtaggtttattttctgttc30
<210>110
<211>22
<212>dna
<213>artificialsequence
<400>110
caagaggaagtccctggggtct22
<210>111
<211>24
<212>dna
<213>artificialsequence
<400>111
tcagatccacttcttctttgccaa24
<210>112
<211>24
<212>dna
<213>artificialsequence
<400>112
agtggagaggaagaactcggaggt24
<210>113
<211>24
<212>dna
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<400>113
tttatgctgtcccaggtactgtgc24
<210>114
<211>28
<212>dna
<213>artificialsequence
<400>114
attggctgatactaacttggaggatttg28
<210>115
<211>20
<212>dna
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<400>115
cccggagaagaaggcaactg20
<210>116
<211>22
<212>dna
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<400>116
gtgctcctaacaaggtggctgg22
<210>117
<211>24
<212>dna
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gctaggggttgcagcctagcttaa24
<210>118
<211>20
<212>dna
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<400>118
ggccttggacagttcccaga20
<210>119
<211>20
<212>dna
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tctcgcagaggttccagagg20
<210>120
<211>20
<212>dna
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agaggtggctcaggctttcc20
<210>121
<211>22
<212>dna
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<400>121
tttgacctcagttccccctcct22
<210>122
<211>20
<212>dna
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<400>122
acccagtaggggcaggcaag20
<210>123
<211>21
<212>dna
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<400>123
agggggaactgaggtcaaagg21
<210>124
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<212>dna
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<400>124
ttgtccctctttgccttctctaa23
<210>125
<211>25
<212>dna
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<400>125
cccaccctctctcatagttttcttt25
<210>126
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<212>dna
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<400>126
atttcccagctgctcacacc20
<210>127
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<212>dna
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ctgttcctatccctggtgcat21
<210>128
<211>20
<212>dna
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<400>128
cctggagctccttcgaatcc20
<210>129
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<212>dna
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<400>129
actgaggcacacagagatgacaag24
<210>130
<211>20
<212>dna
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<400>130
agggtgaggtcagagggtca20
<210>131
<211>24
<212>dna
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<400>131
ggacaggaggactatctacccacc24
<210>132
<211>18
<212>dna
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<400>132
agcctgttccagcctgca18
<210>133
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<212>dna
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<400>133
atggccaagactgcctgttc20
<210>134
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<212>dna
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<400>134
tctagagttgggcccggtct20
<210>135
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<212>dna
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<400>135
aaaacaacacaaaacaggagctgc24
<210>136
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<212>dna
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<400>136
gagtcagggtggatgggatg20
<210>137
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<212>dna
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<400>137
cataccccaatcccaaaggtaaac24
<210>138
<211>22
<212>dna
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<400>138
ccagacacagagccaggtacaa22
<210>139
<211>22
<212>dna
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<400>139
caactttgcagggagagaggaa22
<210>140
<211>20
<212>dna
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<400>140
gaggctgttgggcttgaggt20
<210>141
<211>18
<212>dna
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<400>141
cccaccatgggctgagaa18
<210>142
<211>18
<212>dna
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<400>142
tgccaggccccatgaagc18
<210>143
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<212>dna
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<400>143
tcactctgctgcctccttgc20
<210>144
<211>20
<212>dna
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<400>144
cggcagaagttctccaccag20
<210>145
<211>18
<212>dna
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<400>145
gccctgagcaagcaccag18
<210>146
<211>20
<212>dna
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<400>146
ctgcaactcaggccccctac20
<210>147
<211>22
<212>dna
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<400>147
atctaggggatgggtgaggaat22
<210>148
<211>18
<212>dna
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<400>148
tgcagagcaagccaagca18
<210>149
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<212>dna
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aggacttgcccctcactgct20
<210>150
<211>20
<212>dna
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tcctggatgctcagctgttg20
<210>151
<211>22
<212>dna
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<400>151
cctcatcctcagctcctaatgc22
<210>152
<211>22
<212>dna
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<400>152
gatcgctttggaggactcagac22
<210>153
<211>22
<212>dna
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<400>153
gaacctgcagcttctccatttc22
<210>154
<211>22
<212>dna
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<400>154
cccctcagctaacaagcatctg22
<210>155
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tgggggttggctctcactag20
<210>156
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<212>dna
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agcttgggccccactgtt18
<210>157
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<212>dna
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ctgggctatgagctatgctcct22
<210>158
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<212>dna
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tgttaaaggggctctgaagaagaa24
<210>159
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<212>dna
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atcttctgggggtggggaga20
<210>160
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<212>dna
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tggttccaggagcccagaat20
<210>161
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<212>dna
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gtgaccttgaacttgactctattgga26
<210>162
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<212>dna
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<400>162
tgagagtcacttttattgggaaccatag28
<210>163
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<212>dna
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<400>163
tctcatggggtgaaggctgt20
<210>164
<211>20
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<400>164
ttcctgagcccagagagctg20
<210>165
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<212>dna
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ggagagtagggggccactca20
<210>166
<211>20
<212>dna
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tcccgagtgctcggactacc20
<210>167
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<212>dna
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<400>167
ctgtgggaggccatggtg18
<210>168
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<212>dna
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<400>168
ttcgtgctcattttgtagtttcca24
<210>169
<211>28
<212>dna
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cttagttttctcttctggaaaaatgatg28
<210>170
<211>20
<212>dna
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ctgattggagccgagacctg20
<210>171
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<212>dna
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gtggggccagagtggatg18
<210>172
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<212>dna
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ggggcacttacgactttccc20
<210>173
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<212>dna
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<400>173
cagtactcggccacaccatg20
<210>174
<211>18
<212>dna
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<400>174
gagccacagctggccttg18
<210>175
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gtgaccagagcttttaaccctgtc24
<210>176
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<212>dna
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gctgtggcctgagctcctta20
<210>177
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<212>dna
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ttggtataaaatgtctctgttttccctaat30
<210>178
<211>22
<212>dna
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ttatggaaacaccctgagggaa22
<210>179
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<212>dna
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gaaacagcttgcagactccagttt24
<210>180
<211>22
<212>dna
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<400>180
ttcaggggcatctgatctgaag22
<210>181
<211>20
<212>dna
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<400>181
cccagcctccatttctccag20
<210>182
<211>19
<212>dna
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<400>182
gtggctgtggcccaacgtg19
<210>183
<211>21
<212>dna
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<400>183
ggctatggggagcctctctct21
<210>184
<211>21
<212>dna
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<400>184
cgatcctggtgaggttgttgt21
<210>185
<211>20
<212>dna
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<400>185
aaccagacgcagcctgagag20
<210>186
<211>18
<212>dna
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<400>186
gctcacagcctccttggc18
<210>187
<211>20
<212>dna
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<400>187
caggcaacacctgtccacct20
<210>188
<211>26
<212>dna
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<400>188
tagtggtttctgaaaagcagacagtg26
<210>189
<211>18
<212>dna
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<400>189
tgtccctggctgagctga18
<210>190
<211>18
<212>dna
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<400>190
gggtcttgagccggagca18
<210>191
<211>26
<212>dna
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<400>191
caaaggagacctatgacttcgacatc26
<210>192
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<212>dna
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<400>192
agaacatgttctgggtgatgatga24
<210>193
<211>20
<212>dna
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<400>193
ggctcaagatgctggaggtg20
<210>194
<211>22
<212>dna
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<400>194
tggttttcatggacctgtcgat22
<210>195
<211>18
<212>dna
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<400>195
agctggggagagggctgt18
<210>196
<211>22
<212>dna
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<400>196
gagacaaaccaggccttgagtg22
<210>197
<211>21
<212>dna
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<400>197
gggatgctatgtgccattgtg21
<210>198
<211>20
<212>dna
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<400>198
ggggacatttttgcccagag20
<210>199
<211>27
<212>dna
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<400>199
aatatggaatattacctgttgtgctga27
<210>200
<211>23
<212>dna
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gcctggagttcgagttaatccat23
<210>201
<211>20
<212>dna
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<400>201
gagggaggcctcagtgctga20
<210>202
<211>20
<212>dna
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<400>202
cctgtggagggccttgtgtc20
<210>203
<211>18
<212>dna
<213>artificialsequence
<400>203
ggtgcaggcagagcagca18
<210>204
<211>18
<212>dna
<213>artificialsequence
<400>204
cccctcacctcacccagg18
<210>205
<211>20
<212>dna
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<400>205
ccctccaccaaggtgagctc20
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gagcctcatcctctggacccat22
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cgggcgtcgatccctgtttg20
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gcgcacgggtgctccaagg19
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cccgccttgtgtcgcagacg20
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gcgggcagctggtgctggtg20
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gcgcggcaccagcaccag18
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cagcgtgattcctgggcgatgt22
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gctggaggaggaccaagaca20
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agctgccaggatggactcag20
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tggactggaggctgtcagga20
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ctgaacagcctgcctggtcc20
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gtgccactggggagaggc18
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gggaggcagatgggcactat20
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gacatcgcactgctgcacct20
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agcatgttctcagacaccatgttg24
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gccaagagaaaccgcacctt20
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gaggtagcggctgactttggtg22
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ggctgtgggctccttcacaa20
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aagagcccaaaagagggatgga22
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tgttgataacaagtacaataagcatagaaaa31
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tttatcacacattaatttattcctcttcc29
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gctatcttaaacactagtcaattaatgggaag32
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caatacgaaaacctgaaatccactatca28
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ttttaactgttgtacgcttcatgagtcc28
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gttagcatgataatagtttcctggaagcaa30
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cacaaaagaatattgctttctgacattttt30
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gccttagttttaaaaggatgcctataaatg30
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ctctattgaaatgaagtggcttcatggt28
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tgccttacataaatgcttcaaatttacaga30
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cattattacagtgttgatggagattacatg30
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ggtcttgagtaataagggttccc23
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tccctccacaatggaaaactatgg24
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tgtatgggttctgtatttcagagatgatt29
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ggcaggggcaagattaagca20
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tgtgtttgcccataattctgaatactaca29
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aggctggataatagtcaaattcttacctcg30
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cctgcttttcccccgagaaa20
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tgtttaggtcactgcaactctgactc26
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ttgacagtgggtgaaacaaatga23
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tggtgtaagctgtttccttcatctgt26
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cagatcaagaaactggcgattgaa24
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tttttgcagtttgtttcttcttcaa25
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acatgctgggtcctgcctta20
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ggcaccatctctgattttcaaagc24
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agtgaatttggatatctcgcagagga26
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caggcacaggctcgcgatgg20
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cggcttctgctccggggtgt20
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gccgtggggctcgtgcattc20
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atgccagctaaggtgctttggtagca26
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ctgcgagtaccagtgccagc20
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ctcgaaggtaccggggaggt20
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cgtcctcgcaccggtgttggt21
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cggagcggtccaggggcact20
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agaccggctaccggctgg18
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gcgcagacgcagaggtagct20
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ctcgcagccgccgttctcca20
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gaggagcagcagtgcgaagtgaagg25
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ggcctgcaggtggctgggaag21
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gccggcgcctctggatcg18
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aggggcccgaggcgcttg18
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ggttccccgcacccgcagag20
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cccgggtagagcgcgaagca20
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cgcgcagcggcaagaagtgt20
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aggggcacaggacgccgatg20
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gcccttcccaccaggcacttcc22
<210>271
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acgttcgggaaaaggaagga20
<210>272
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aagagatgaaagagggctgcac22
<210>273
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cgcaagggcggccagagaac20
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gccggagcagaggggcacag20
<210>275
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cagcacccctgtaacaagacg21
<210>276
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cctgtcttcggggaagtcca20
<210>277
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cctgctaaattgatttcattgtcca25
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tcatggtggtgacagatccaa21
<210>279
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ttacttaaaattctggtaaaatgtagccatta32
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ttctccttgggctgcttcct20
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cggcaggacttctcaccaag20
<210>282
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atggaaggaggccaggagat20
<210>283
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ggcaggtctgggcaggag18
<210>284
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gctggctgcagtctagggaaaa22
<210>285
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ccatcctcctctgcccagac20
<210>286
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tgagattctccagaaccatcctga24
<210>287
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ggggttattatcttccgctgcc22
<210>288
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ctgcaagggctgcagctg18
<210>289
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gcacgtgctggaaggcccaga21
<210>290
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ccccactagccccgcccaga20
<210>291
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tggcaccctctctgcacagt20
<210>292
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tactgcacacaggtgtcctcca22
<210>293
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caaccgcactcggtatgaactg22
<210>294
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ttgagtcaaaccccaaagcgtt22
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gggcctattcttcgggctaact22
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ttctggtgtttcagttggggagtc24
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tggcaacaaacacagcaacc20
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ccaggctgcattgagggtag20
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gttggaggcagatgcatggt20
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gccaaggtcatcagtgggatt21
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caacagcccagcaccaaagttc22
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gaaaaatgaaaccatttcccacttctaa28
<210>303
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ctgccatgtggatgctgcag20
<210>304
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ttttccttgtggaagtcggcag22
<210>305
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aaataacaaaaacctgagcatgcctc26
<210>306
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agacgctggatccggctctt20
<210>307
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gctgggaacatcctccagcttt22
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tgatttcatacttgctgctggcat24
<210>309
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gaagggagagacccatgaacaagt24
<210>310
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ggcagggtctgagaagtcagct22
<210>311
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agtttccaatcctgcttgacttca24
<210>312
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acaaaaatagtatgttgtgggtttgctg28
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gaggcctccagggaaatcag20
<210>314
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tgtgtgggaccacaattagca21
<210>315
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ggactgcgacatcctccagc20
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attaacagagtggccctgtatggaag26
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ttgtaacttgtggttcaataaaactggg28
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gctgtgctgaggcagaccct20
<210>319
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gctgacctccagaatctgacaacttt26
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gcacttcagacacctagacctcca24
<210>321
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gatgaatgccatcagtccctcc22
<210>322
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gtactgcacagaccctccccag22
<210>323
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caagcctgccaagaggtatca21
<210>324
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ggggctaggggtaattttcct21
<210>325
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tccactgactgatgatggagattt24
<210>326
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acaaaaccttgatacatttttgttgtg27
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gaacccagggtcaaggtgaaca22
<210>328
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gccttttattgatcaggtctagagctttct30
<210>329
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tgcagcactgcttttgatcaca22
<210>330
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ctctgaccaaagccattcattgtg24
<210>331
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tccatgtgctactcacatgttgct24
<210>332
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tcttctactgtgttagcccaggca24
<210>333
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gatgcaatgactcagcaagtcctc24
<210>334
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ggagtctatgaaggcttttctttcatgt28
<210>335
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ccaagccaccattaacatttccag24
<210>336
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tggagcctgggccttcag18
<210>337
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tggtgattttgaattctcatactgc25
<210>338
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tgactctagtcaacgatcacagaca25
<210>339
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cttcacaccacctggacacaca22
<210>340
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actcatagagctttaggaagcaacaacact30
<210>341
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gttacatgatgataggcacttttctgtgac30
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tggcatgttgacaacttgagca22
<210>343
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cacttccacaaggccctcct20
<210>344
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atcatggcagtggggatggt20
<210>345
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tcatgaacatgatacccatagacag25
<210>346
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ggcagcacctcaccttttctt21
<210>347
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ccttccattgcagacaaattgg22
<210>348
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ttcattttcctcttcaaaggaatcacat28
<210>349
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cattgccgcaccacaaacat20
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ttgatctctcttctcaggccatctc25
<210>351
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agacaaatctatgccactcagaatctcc28
<210>352
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gaagcaggagagcattcggaaa22
<210>353
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cacacaaggaaaaggtattataagcagaga30
<210>354
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ttggttggaaacaggaagtctgaa24
<210>355
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tggattttatgttcccttctgatgc25
<210>356
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ttccgatcaacaatctcatctgg23
<210>357
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tggaaggccacccaggtatg20
<210>358
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aagcagcattactcttactccttgctta28
<210>359
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gtgctggagattgtgccaaa20
<210>360
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<212>dna
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gcattgatcatgcttctgtttca23
<210>361
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<212>dna
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gactagagtgggcacacagtagatactca29
<210>362
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<212>dna
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cgaagctggaatttgatgaagca23
<210>363
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<212>dna
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ctcataatcgagccactgtttaagttt27
<210>364
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tccacagtaaccaggcaatgct22
<210>365
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tccatgatcatttcaagttgtcacac26
<210>366
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gcattaaaatagctaaagaagctgtgatgg30
<210>367
<211>22
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gcaatccattttccggcttaaa22
<210>368
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<212>dna
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caccctaaatgcctttttatcacattt27
<210>369
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<212>dna
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<400>369
tgcattgagctttctgtatttcttactctt30
<210>370
<211>26
<212>dna
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<400>370
gcatcaaagtgggaagatattgatga26
<210>371
<211>22
<212>dna
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caaaccttttcggccacttttc22
<210>372
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<212>dna
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caaaaatgcatgacctcacacaaa24
<210>373
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<212>dna
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tcctctgcctataagattgaacatttaggg30
<210>374
<211>30
<212>dna
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<400>374
ggatctctcattttagaaaacaggtgagaa30
<210>375
<211>22
<212>dna
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tgatgtcaaacaaagccaatgc22
<210>376
<211>24
<212>dna
<213>artificialsequence
<400>376
ccaggaaacacaaatcaagggttc24
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tttgctagtaaaatttctctaactgggatt30
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cccccagctcatcaggattg20
<210>379
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ccttcagccattccagaccagt22
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agggatctttgcattcatttatgtctaaaa30
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ttgcctaataggctcactttttaatttagg30
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atccatcttcattgcatggcag22
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atggaaacatttagtgttgcattttgaa28
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ttttgttttgttttttcaattgatggtaga30
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aaatgaaacctcactcccaagga23
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tttaggtttgctaagatatgttttcttttt30
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ggctggaaatgttcagtctgtag23
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tgtcatacaattcataggcagaaaatg27
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cagttcttcgatgcattctctttcaa26
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caccaaacccgctctctgaa20
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tcgacaatgctggggaagggagaa24
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ctgggagcgggcggtctcct20
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cagcgagcctcggcggaacag21
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gggatcctgtcctcttgaaccctgga26
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gtggggaacgcagttctctg20
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atattcccttcttttcttcatgttcttta29
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aaagcaaatggggtcattttga22
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aaattgcgccactgttctcc20
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ttgctccgataaagacttcagttg24
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ggcaggcagatcactcacg19
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acttgaacctggaaggcgga20
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gcctaagcagataggattcctcct24
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aagaagcaagcaagctgaaatagc24
<210>404
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aaaagaaaacatggaaagtctaatgga27
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gggaaactgactgggcaaga20
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attttgcctgtgtgcctgct20
<210>407
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tagactttgagattcaaggtcagctaga28
<210>408
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tagttctcaatgttatgtaacattctctctca32
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agattttaacgactttcacaaatatgctgt30
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cttgagaggaatgttaaatagtaagttctgaa32
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tacattttatgttctaaaagcatgcacc28
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tttttcacctcaattcctaccctg24
<210>413
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agtaaaatccaacataacgcatgc24
<210>414
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atgttagatggaaatgtgttaagaatagcaat32
<210>415
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ccattattttaaaaacataaattcctattcat32
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catcacagctggtatttgttgattct26
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agaatctggaaggtactcatgtcttctg28
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aaagttgatggcaaaagctttttagtaa28
<210>419
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<212>dna
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gttggaagaataagacacttttcctttttc30
<210>420
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ctgagtgagatctactgtaatcatcagcatat32
<210>421
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ctggtgaattgagtccctgaca22
<210>422
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<212>dna
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ggattttgatcagctatttttagaagataaca32
<210>423
<211>26
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tgctagcatgagctgactttactttc26
<210>424
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gctggtcactctcagccagaa21
<210>425
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ccgaggaggctgatacatgc20
<210>426
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agacgggacaagggtgtctg20
<210>427
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agacacccttgtcccgtctg20
<210>428
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gaatgttctcccttctgtggcta23
<210>429
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gaagatgtaggaagctgctcatca24
<210>430
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aatgtgtgaagaagatgaactaataaaaacag32
<210>431
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tttttagtaaaggaaatttctttccctctg30
<210>432
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ctagtagtgttaaacattttatccagttcacc32
<210>433
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caaagaaaacacagataatgtacagtgg28
<210>434
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catgacttgacagggcagaaa21
<210>435
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tgcatatatgtttatgtgtattgtgtatgg30
<210>436
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cccaacgcattaagcattcc20
<210>437
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aggcaaaatcagcctgagca20
<210>438
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gggtccttcagggactccaa20
<210>439
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gggattttctgtttcgtgcaa21
<210>440
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cttgggagactgaggcagga20
<210>441
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ctggatccgggagcacac18
<210>442
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agctgtgaatcgcttttctgtctt24
<210>443
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ggccgtcaggcacaaatc18
<210>444
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cactgcggaatcaccaagga20
<210>445
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ttgtcagatggggtgtgaagaa22
<210>446
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gtcggaaacacgcagctcag20
<210>447
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tcgcgggcttcttccgccta20
<210>448
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cccgcctcacgctcctctcc20
<210>449
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cgggcggagaggagcgtgag20
<210>450
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cagggggcttccccagaacg20
<210>451
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cagccgccttccctgaccagga22
<210>452
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gcgctgggagacggaggagc20
<210>453
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<212>dna
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cagccggcaccgccgaag18
<210>454
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actcttgcggagccgctgcc20
<210>455
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cacgacggggcgccgttaga20
<210>456
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<212>dna
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tgcccagggagccccctttc20
<210>457
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cgggctcccggcgctcta18
<210>458
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ctgggactgaggcggggtcc20
<210>459
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ccttctgcttgccccagac19
<210>460
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ggaggagcgtcaggaagaca20
<210>461
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<212>dna
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ggtcctagactctcctgagaccacta26
<210>462
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tgtcttcctgacgctcctcc20
<210>463
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tctgtaggcccagggttgc19
<210>464
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aggttcaagaagggccttgg20
<210>465
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gccctcagccctggtaagac20
<210>466
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gagagagccccaggccac18
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gagggacagtgcctgagcag20
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ggcagggtacatgtctccca20
<210>469
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gggagatgggccttaagaagac22
<210>470
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accatacacatccccaccca20
<210>471
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cccagaacaatcctgggaca20
<210>472
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cccacaaactcccaactttcc21
<210>473
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gtggaaagccaagtggagca20
<210>474
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aagtgggtattgttgtaagatgctga26
<210>475
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aaatgaacacacttcacagtggactc26
<210>476
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gtttgtggaaagccaagtggag22
<210>477
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tggtggtgggctcctgtagt20
<210>478
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ggggatcttggagtccactg20
<210>479
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ttcactgggaataaataaattttgaaga28
<210>480
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ggaaccttccatgacgagacc21
<210>481
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aacagccctgcctgttgg18
<210>482
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gctgggaagtggtgaaagca20
<210>483
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acatgacggcagcgtggag19
<210>484
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ccagcagtgcccagaaagtg20
<210>485
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tttgaaccctgctgagccagt21
<210>486
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tcaaaaattaaatgaattgcacataaagc29
<210>487
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tgacattgatttactgaccatttattccac30
<210>488
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ttacagattcctgcgtaggtgcaa24
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ccaaataactgaaagaggcagcaat25
<210>490
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gggaggatggggaggacagt20
<210>491
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ttactgcagcctttttgcagatg23
<210>492
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<400>492
ggtctgtggggagaagtggaa21
<210>493
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ccactgtgtggacttcccttca22
<210>494
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gcaaatactgcaccattttatataagggact31
<210>495
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gagcatctccttctgccttgc21
<210>496
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ccctgtgggaaggggcttta20
<210>497
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ttttcttcttcctctctgtccttccttc28
<210>498
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tcaggcgttcacttcttgagga22
<210>499
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ccatgaatgtaaatgttcagtgctactaaa30
<210>500
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ggacttgtaggtcacttttcagaggc26
<210>501
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ctgggcgacagagcgagact20
<210>502
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aaggcaaagtcaatgggatttca23
<210>503
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ctttttggaaagctaaactcacaatcactt30
<210>504
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gggccctcagtaagctcccata22
<210>505
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tttttcagtaattgttaagcttgatttcttt31
<210>506
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tgaagcatgaagcatggctct21
<210>507
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tcccacctcttgtgacctgtattg24
<210>508
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ccatggcttttcatcccagc20
<210>509
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ctgtggctcttcataatcatccattttt28
<210>510
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tgagcaaaaggccaaagagtcc22
<210>511
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gagctggagcttacatgccttc22
<210>512
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gagtggggtctgatggagagg21
<210>513
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caccacaaaattgctaccttgtctca26
<210>514
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ggattttatgtaaatgtagaagggttttgc30
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acaatggagcagaacaaagtatcaatttaa30
<210>516
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cccttcccttctctcttatccaattg26
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aggggctggaccatattttcct22
<210>518
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atggcccataaaacccagacataa24
<210>519
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ctgcgagcagagcagtcaaa20
<210>520
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gcccaggcaaaatcaggagt20
<210>521
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tgggtgcagttgccatttcttt22
<210>522
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cctggggaatgaagtctgaggg22
<210>523
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gcaggcagcctaaccctcac20
<210>524
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ccagcatgctaaatccctaccc22
<210>525
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ccattcaaaatacaatttaacaatgcaaca30
<210>526
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aatccttcaaaatggggaacaaca24
<210>527
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atactacatatacaacatgtgtctgtctagca32
<210>528
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gacacacctacacttcctgcca22
<210>529
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agagagctgggaagggatcc20
<210>530
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cgcctgatgaggagagtcgc20
<210>531
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tgctgagatctggtgccg18
<210>532
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caccagacagctctccggat20
<210>533
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<212>dna
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tgtgggtttctggtccagtg20
<210>534
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ggtcagtatggaatgggacagg22
<210>535
<211>18
<212>dna
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cccgcccagtagtgagca18
<210>536
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<212>dna
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<400>536
ggtagctcagaataccccaccc22
<210>537
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acaggccagagccactatgc20
<210>538
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<400>538
ccaccaaaaatcccagacca20
<210>539
<211>22
<212>dna
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ctctagaaccatcgccctctgc22
<210>540
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ctgcacagtccagcgtgg18
<210>541
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gcccacccgacgggttac18
<210>542
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<212>dna
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catccgggacaagaggttaaag22
<210>543
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<212>dna
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cctggttggggtgtcctaaa20
<210>544
<211>18
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gagcagccttcccaggct18
<210>545
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<212>dna
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ctcagcctcctgccctctg19
<210>546
<211>20
<212>dna
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gggtacccaggtcagctcct20
<210>547
<211>20
<212>dna
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gtggggacggacgtagagagaggtg25
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ccaccgccatccccctcctc20
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gctatgtcgtggctgggaag20
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caaacgagcaagcagccata20
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gcacctgccaccccatcacc20
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cttcccctgggtggcaga18
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gacggggatcgctgggtcctc21
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tgagtgctggtccccaggagcat23
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gctgagcctgtcccttgg18
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catctgcacacaccaggagc20
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ccttctggcagggtcagctgtga23
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gaccaagtgcttgtgccagtc21
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aatcggggaagcactgctta20
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ctctgctgaccaggcgtg18
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accggtctgtctgggttcct20
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acagcacaagtccccatgct20
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attacttgtggccggtccttct22
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aacaaatgttcaatgaccctcagg24
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ccctagccttttggacattgg21
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ctttagagtcttctccaatgtccaaaag28
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cggagtggaatcctgattgg20
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caaaaacaaataaactggactayatcaaaa30
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caccgtatctttgtcagagagcaa24
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ccactcatacattgctgatggaa23
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tccctcaatgggtctttgga20
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cccaagaggccattggaaat20
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tgcatctgaagggtattatgtgga24
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ggtgctttgagtgatgttatccaa24
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tttttctagatcaaatgtattatgcagtaaga32
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cattgccaattaggtcagtggtc23
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tgttcgtgtattccttgtcagca23
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atctctacctccttcatggaagcc24
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ggctatgtaagtggctggggaag23
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ccatatttgcctttcattgcaca23
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caattccaatgaattaaccttggaaa26
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cagcaggctcaaaggcataa20
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ctggagaatgggggtgctg19
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ccatagcaatcatgcgagtcaa22
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cagaaggcagaggagcctga20
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aacaggagccgccagaaaac20
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tagctcacagcatggccttatg22
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gaggggaaggaggtgatgttg21
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gtgggcgcttcacggaactcg21
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cacagccgcagccaaagagacg22
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ccagggccctctgcaaat18
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ccacccgtcttttgtccagtaa22
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ggagggcgagtcatcagaga20
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caggttcctccagttcttgattttg25
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gcaggtggtggaaagggcctga22
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caccttcctccaagcagacttaca24
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gggcaaaagcagagagactaacag24
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gaactgaagtagcaaaagggaggc24
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cagcagcaagagaagttctgga22
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cttccctggtggaagccata20
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gaatgggcaccactcctgaa20
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ggcgatggttcctcacaaga20
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ttgtccattctgtcctagcaagtg24
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tgagtatggggaagagagagtacca25
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ctgtccagaggaaccccaac20
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ggcaactgagcgaatttgga20
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ccaaatattttgagttttatctagcctcaa30
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ggcatatattcaggcctgacactt24
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ctcctggcagaattggaaaatc22
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gcatttctttacccctttcaatatatgtaa30
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ttttgcaggcatcaatggctat22
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gcattcacagctgttggtacaaga24
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gcaaacaaactgacaactgcaaac24
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cttcagtcagatcaagaggaaattga26
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gtcacaagagcagagcaaaggaat24
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gaaatcccaagagaagtcaccaga24
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tcttcatttcaactgatatggtatcatcat30
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tccctacggaaactagcaatgg22
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gaaagcagtcatttcttacaaggagc26
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tcccttccacgagatccaga20
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gagaaggagaaaggggccatta22
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ccaaggttagaatggctaaagaaagg26
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gcaccacaaggatatctcctaatacaag28
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tcgtaaggcaatctgataagggag24
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tcttactgagcactaggcaaaatgtaga28
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<400>718
tgggagtctgaattgtttcaaagc24
<210>719
<211>24
<212>dna
<213>artificialsequence
<400>719
ggactcaaagagatggtttttcca24
<210>720
<211>30
<212>dna
<213>artificialsequence
<400>720
taatgacctaaaatcttgaagtactggagc30
<210>721
<211>22
<212>dna
<213>artificialsequence
<400>721
caccagatgcacaaaatccaga22
<210>722
<211>30
<212>dna
<213>artificialsequence
<400>722
caagttagtaagaaataggtttctgctgct30
<210>723
<211>22
<212>dna
<213>artificialsequence
<400>723
aagactcacattgatggcccat22
<210>724
<211>28
<212>dna
<213>artificialsequence
<400>724
gattctggcaagaatagcatcttaaaga28
<210>725
<211>30
<212>dna
<213>artificialsequence
<400>725
agttgttagaatcaggtttaatgaatagcc30
<210>726
<211>24
<212>dna
<213>artificialsequence
<400>726
tgccagactgatggactattctca24
<210>727
<211>30
<212>dna
<213>artificialsequence
<400>727
tacatcaaataatctgatttcaacaattcc30
<210>728
<211>28
<212>dna
<213>artificialsequence
<400>728
taaataacctaccactctctgttgacga28
<210>729
<211>26
<212>dna
<213>artificialsequence
<400>729
tctccaagaagccaaatatgagactt26
<210>730
<211>26
<212>dna
<213>artificialsequence
<400>730
cttgtattatcagtacctgctgccaa26
<210>731
<211>24
<212>dna
<213>artificialsequence
<400>731
caagaagcttctcccagaattcaa24
<210>732
<211>24
<212>dna
<213>artificialsequence
<400>732
ctattgctccaggtgatggatcat24
<210>733
<211>28
<212>dna
<213>artificialsequence
<400>733
tgagtaaccagagtcttattcttcctca28
<210>734
<211>22
<212>dna
<213>artificialsequence
<400>734
tggcattaaattgcttttgcct22
<210>735
<211>30
<212>dna
<213>artificialsequence
<400>735
cacttggtacataaaattattgcttttcct30
<210>736
<211>30
<212>dna
<213>artificialsequence
<400>736
aaaattaattttcttgtaattccactgtcc30
<210>737
<211>22
<212>dna
<213>artificialsequence
<400>737
gaggcaaggagcagaacctaga22
<210>738
<211>30
<212>dna
<213>artificialsequence
<400>738
caaaattcttagtacacaaagaccatttct30
<210>739
<211>28
<212>dna
<213>artificialsequence
<400>739
ccaaaagacaggtatttcttttagggat28
<210>740
<211>26
<212>dna
<213>artificialsequence
<400>740
aagttttggtttcattaggcttgaca26
<210>741
<211>24
<212>dna
<213>artificialsequence
<400>741
aggaatttgctctgtttttcacca24
<210>742
<211>24
<212>dna
<213>artificialsequence
<400>742
tcaagtccctggatcaagtctcat24
<210>743
<211>30
<212>dna
<213>artificialsequence
<400>743
ggtgagagagcttatactcataggattgat30
<210>744
<211>26
<212>dna
<213>artificialsequence
<400>744
attttcagtgaagtaccagcttttgg26
<210>745
<211>28
<212>dna
<213>artificialsequence
<400>745
tagattttaacaggcttctctgtgtcct28
<210>746
<211>30
<212>dna
<213>artificialsequence
<400>746
caaatagagtaggtagaagaaagagcacaa30
<210>747
<211>28
<212>dna
<213>artificialsequence
<400>747
aagaaataatttctgttcctgttggttt28
<210>748
<211>30
<212>dna
<213>artificialsequence
<400>748
aagctgtaaagaagtaggctgagtaggtag30
<210>749
<211>26
<212>dna
<213>artificialsequence
<400>749
aaaaatagcatttgttgacgttctcc26
<210>750
<211>22
<212>dna
<213>artificialsequence
<400>750
taatcagcccaggttcttggag22
<210>751
<211>26
<212>dna
<213>artificialsequence
<400>751
ttgatactggtatgccataaagcctt26
<210>752
<211>28
<212>dna
<213>artificialsequence
<400>752
aatgtgatacatttcccatcattgatta28
<210>753
<211>30
<212>dna
<213>artificialsequence
<400>753
caggaggtagcacatacatttaaaaatagg30
<210>754
<211>28
<212>dna
<213>artificialsequence
<400>754
taaagtattcaggcattccctttaaatg28
<210>755
<211>30
<212>dna
<213>artificialsequence
<400>755
gctttttactctgtattcactttccatgta30
<210>756
<211>24
<212>dna
<213>artificialsequence
<400>756
atggttgagggaagaaggatatgg24
<210>757
<211>26
<212>dna
<213>artificialsequence
<400>757
agcatgtccttgtgataaccttcttt26
<210>758
<211>30
<212>dna
<213>artificialsequence
<400>758
cactgctctgagtcagttaaacagtaaatc30
<210>759
<211>26
<212>dna
<213>artificialsequence
<400>759
ttctttctttctttctttccaaggag26
<210>760
<211>24
<212>dna
<213>artificialsequence
<400>760
ggtgaccaagaggctactagtcca24
<210>761
<211>23
<212>dna
<213>artificialsequence
<400>761
catcttgctattcagtgccccta23
<210>762
<211>22
<212>dna
<213>artificialsequence
<400>762
gaggagttaaccccaaaggtga22
<210>763
<211>22
<212>dna
<213>artificialsequence
<400>763
tttgcagtgaccattgtcctgt22
<210>764
<211>20
<212>dna
<213>artificialsequence
<400>764
gagctgaggagggagaggtg20
<210>765
<211>22
<212>dna
<213>artificialsequence
<400>765
ccacatcacaggccatgtaaga22
<210>766
<211>22
<212>dna
<213>artificialsequence
<400>766
tgggaagcacccatggatatta22
<210>767
<211>23
<212>dna
<213>artificialsequence
<400>767
ttgcagtgaactgtgatcccatc23
<210>768
<211>20
<212>dna
<213>artificialsequence
<400>768
cccctgctgtggaagtcacc20
<210>769
<211>23
<212>dna
<213>artificialsequence
<400>769
tctcctagagggcaaaagccata23
<210>770
<211>23
<212>dna
<213>artificialsequence
<400>770
gagcaaaagaccccatgttgaaa23
<210>771
<211>25
<212>dna
<213>artificialsequence
<400>771
caaggaaaatacctgggaatttgaa25
<210>772
<211>27
<212>dna
<213>artificialsequence
<400>772
tgaggagataacatttcaactttgttc27
<210>773
<211>20
<212>dna
<213>artificialsequence
<400>773
agaggtgggggcatctccag20
<210>774
<211>20
<212>dna
<213>artificialsequence
<400>774
ggtgtcagggtgccaaggag20