CN113981102B - 一种基于二代测序技术检测y-snp单倍群的引物组合物、试剂盒和方法及其应用 - Google Patents
一种基于二代测序技术检测y-snp单倍群的引物组合物、试剂盒和方法及其应用 Download PDFInfo
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Abstract
本发明涉及生物技术与遗传学技术领域，提供了一种基于二代测序技术检测Y‑SNP单倍群的引物组合物、试剂盒和方法及其应用。本发明依据中国人群单倍群频率，参照最新版2019年ISOGG网站(Version:15.58)筛选出了381个Y‑SNP位点，利用二代测序平台和多重PCR扩增子捕获技术，设计了一个针对中国人群单倍群检测的方法。该方法灵敏度高、特异性好，对案件检材适用性较强，对降解检材及混合物检材也表现出良好的检测能力，可获得高分辨率的单倍群结果，单倍群划分细致、推断准确。所述方法可应用于家系排查和对未知样本地理来源的父系族源推断，为法医学实践提供一种灵敏度高、分辨率高且系统效能强的族源推断技术。
Description
技术领域
本发明涉及生物技术与遗传学技术领域，具体涉及一种基于二代测序技术高分辨率检测Y-SNP单倍群的引物组合物、试剂盒和方法及其应用，用于扩增381个针对中国人群主流单倍群的Y-SNP位点。
背景技术
Y-STR检验是一种非常成熟的法医DNA分析技术，在辅助刑事案件侦查等方面也有着极为广泛的应用以及独特的应用价值。但是由于Y-STR突变率高，表现出同一家系出现单倍型不一致的问题，这对家系排查带来了困扰，阻碍了对样本来源的推断。因此，解决Y-STR突变造成的潜在混淆，正确判断犯罪现场未知检材的地理、民族甚至是家系来源，是法医遗传学亟需解决的科学问题。
Y-SNP是另一类重要的遗传标记。SNP突变率低，不会发生回复突变，遗传稳定性强，且适用于降解检材。这些特性使得Y-SNP成为研究进化和迁移的理想标记。由多个Y-SNP构成一个单倍型，而一个单倍型因突变的发生会衍生出多个新的单倍型，这些源于同一祖先的单倍型被合称为一个单倍群(haplogroup)。如果所有男性都来自同一个父系祖先，那么这个家族理论上都属于一个Y-SNP单倍群。单倍群互相关联、合并，形成一个发源于共同祖先(the most recent common ancest,TMRCA)的Y染色体进化树(Y ChromosomeHaplogroup Tree)。2005年，国际遗传谱系学会(ISOGG)成立，并于2006年在ISOGG网站上发布了Y-DNA Haplogroup Tree。发展至2019年，ISOGG网站上(version:14.177,date:8October 2019)提供的可用于构建Y染色体进化树的Y-SNP位点(包括Indel位点)已达69280个。细分之下的Y染色体进化树能追溯到更短世代数内的最近共同祖先，使其能应用于未知男性样本的地理、民族甚至家系溯源。同时运用Y-SNP单倍群也能克服在法医学溯源过程中因Y-STR突变造成的不利影响，即来自同一父系的个体包括亲缘关系较远的个体均拥有完全相同的Y-SNP单倍群分型，可对应于Y染色体进化树上的同一分枝。
近几年国内外学者对Y-SNP单倍群的研究越来越多。例如，Arwin Ral等用850个Y-SNP位点构建了一个世界范围内的Y染色体单倍群树，包含共计640个Y单倍群，囊括了各主要单倍群(A-T)及其亚群。因为该树是针对全世界范围的，数量上针对各个单倍群没有明显的取舍，因此该树针对某一区域分辨率较低，极大地限制了在实际应用中对未知男性检材的具体来源推测。国内的Yin等学者和Zhou等学者亦开发了很对中国人群单倍群的Y-SNP复合扩增体系，但包含Y-SNP数量较少，对人群的区分度依旧不足。因此，需要寻找更多中国人群主流的Y-SNP单倍群下游的枝系，并构建高通量检测体系。
目前用于Y-SNP检测的主流技术有基于多重单碱基延伸的SNaPshot、MALDI-TOFMS和二代测序(next generation sequencing,NGS)等。由于SNaPshot有着较高的灵敏度，以及与毛细管电泳平台的兼容性，在法医学领域有许多基于SNaPshot建立的多重Y-SNP复合检测体系。但其成本相对高、设计和优化体系较难、操作流程复杂，耗时较久。MALDI-TOFMS通过直接检测分子质量差异，准确性和成本上都非常可观。但以上技术通量有限，能检测的Y-SNP数目在50个以内，无法同时对多达几百个Y-SNP位点进行检测。因此一个通量高又适宜在法医领域推广的检测方法非常重要。NGS技术在准确性和通量方面具有巨大优势，适用于多样本、多Y-SNP的检测。将其作为技术手段在构建高分辨率Y染色体进化树时，不会受到检测Y-SNP数量的限制，从而在每一次的检测中都保证了体系的完整性，避免分型错误。
发明内容
为了克服现有技术中的缺陷，并构建一个适用于中国人群的、高分辨率的完整Y染色体进化树，本发明根据各个地区和群体的Y染色体遗传结构特点，基于二代测序技术提供了一个用于Y-SNP单倍群复合扩增检测的引物组合物、试剂盒和方法。
为实现上述目的，本发明采用如下技术方案：
本发明的第一方面是提供一种基于二代测序技术检测Y-SNP单倍群的引物组合物，该引物组合物包括381个Y-SNP位点的扩增引物；
上述381个Y-SNP位点包括AF6、P305、P108、M60、M91、M130、F3393、CTS11043、M8、M356、M38、M217、L1373、F3447、F1699、F3918、FGC28881.2、F1756、F3830、Y10420、M48、M86、B90、M504、F3796、F9992、F1067、Z1338、Z1300、CTS2657、CTS11990、CTS8579、M407、F3850、Z12209、F1319、CTS3385、FGC45548、F845、CTS10923、MF2091、P143、M168、M294、CTS3946、JST021355、M174、CTS11577、M15、N1、Z27269、PH4979、PH1991、Y7820、SK541、Z31625、Z31584、P99、P47、M533、Z34359、PH97、Z42599、Z42600、Z42601、Z42602、M116.1、M145、M96、M89、M427、M201、P287、P15、CTS4367、L30、P303、L140、F1329、L901、M69、M52、M82、M578、M170、M429、M522、M304、M267、M172、M410、L26、M47、M67、M9、M526、M20、M22、M27、M317、P326、P256、M231、F3373、CTS3750、CTS11499、F1206、Tat、M178、L708、P298、L392、CTS10760、Z1936、B197、M2019、F1008、M128、B523、P63、B525、F2930、CTS582、M1819、F549、M214、M175、F265、M119、B384、P203.1、F446、F140、CTS3265、F78、F81、F533、F492、F656、SK1568、Z23406、M101、Z23392、CTS8423、CTS52、CTS701、Z23266、CTS716、CTS352、Z23271、M50、F1036、P31、M268、F2320、M1470、PK4、M95、F1252、M88、F2758、F2890、CTS5854、Z23810、Z23781、F789、F838、CTS1456、F993、F417、CTS9996、M176、P49、M122、M324、F113、L127.1、KL1、L467、F1876、F2159、F1867、F852、F2266、L599、Z43961、Z43963、F854、Z43966、F915、IMS-JST002611、F240、F18、F117、CTS5820、F13、F11、F632、F110、F17、F377、F856、F1095、F1418、F1495、CTS7501、F793、Y20951、Y20932、F38、F12、F196、F930、F2685、F1365、F3232、Y15976、Y26383、FGC54486、CTS12877、F2527、F723、F1422、CTS2154、SK1691、PH203、F449、F238、F134、F1273、F724、F1266、CTS498、FGC3750、P201、M188、F2588、CTS445、CTS201、M159、M7、F2680、F1276、CTS6489、F1275、M113、N5、Z25400、F1863、F1134、F1262、Y26403、F1837、F862、F879、F1226、F2859、P164、M134、F450、M117、M133、F42、F438、Y17728、F155、F813、Y20928、F1754、F2137、F1442、F1123、F1369、F310、F402、F1531、Z25853、CTS5492、CTS6987、Z42620、CTS4658、Z25928、SK1730、Z26010、A9462、CTS7634、F317、F3039、Y29861、CTS5488、CTS10738、CTS9678、Z39663、A9457、YP4864、Z44068、F5524、F5525、Z44071、Z44091、CTS4960、F122、F114、F444、F79、F629、F46、FGC16847、F48、F2505、F242、CTS4266、Z26108、F2887、F3607、F3525、CTS3763、A9472、FGC16863、L1360、FGC16889、SK1768、F4249、FGC23868、CTS335、CTS53、CTS6373、A9473、F3386、Y29828、FGC34973、F1739、F743、F748、AM01822、N6、AM01856、N7、F4110、F4068、F4124、AM01845、F717、A16433、F742、F1150、F837、F1025、F1055、F3021、P295、M45、M242、F903、M346、M3、M207、M173、M420、SRY10831.2、M17、Z645、M458、Z91、Z93、F3105、Y7、Z2124、Z2123、S4576、F1345、M343、L754、L389、P297、M269、M479、M124、FGC21706、B254、M184。
进一步地，上述扩增引物组合物包括核苷酸序列为SEQ ID No.1～746的引物。
本发明的第二方面是提供一种包括上述引物组合物的基于二代测序技术检测Y-SNP单倍群的试剂盒。
进一步地，上述试剂盒还包括多重PCR反应所需的其他试剂、接头序列PCR反应所需的试剂和磁珠纯化所需要的的试剂。
进一步地，上述试剂盒检测的样本来源于血液(血斑)、精液(精斑)、唾液(唾液斑)、骨骼、毛发、人体组织，然后采用血液(血斑)、精液(精斑)、唾液(唾液斑)、骨骼、毛发或人体组织基因组DNA(gDNA)。
本发明的第三方面是提供采用上述试剂盒的基于二代测序技术检测Y-SNP单倍群的方法，包括如下步骤：
步骤一，多重PCR反应：以稀释好的gDNA为模板，并配制扩增反应体系进行多重片段扩增；
步骤二，对PCR扩增产物进行磁珠纯化，然后进行接头序列PCR反应；
步骤三，对步骤二的扩增产物进行磁珠纯化构建成文库；然后对文库进行浓度、片段长度和纯度进行测量；
步骤四，将文库混合、变性后加入Illumina MiSeq测序平台进行测序；将测序结果与参考基因组序列比对，采用samtools、GATK等软件寻找SNP和Indel，然后运用ANNOVAR软件对SNP、Indel位点进行注释、确定突变位点对应的基因信息等。
进一步地，步骤一的多重PCR反应体系为25μL，包括10μL ddH2O、4μL Enhancerbuffer JP(2N)、4μL引物混合物(2μM)、1μL gDNA以及6μL IGT-EM101 polymerasemixture。
进一步地，步骤一的多重PCR反应条件为：预热105℃；95℃3分钟30秒；14个循环：98℃20秒，60℃3分30秒；72℃5分钟。
进一步地，上述接头序列PCR反应体系为25μL，包括17μL PCR磁珠纯化的产物，1μLIGT-I5 Index(10μM)、1μL IGT-I7 Index(10μM)和6μLIGT-EM101 polymerase mixture。
进一步地，上述接头序列PCR反应条件为：预热105℃；95℃3分钟30秒；9个循环：98℃20秒，58℃1分钟，72℃30秒；72℃5分钟。
本发明的第四方面是提供上述引物组合物或者试剂盒在家系排查和对未知样本地理来源的父系族源推断中的应用。
本发明采用以上技术方案，与现有技术相比，具有如下技术效果：
(1)本发明基于二代测序技术构建了一个包含381个Y-SNP的扩增引物的引物组合物和试剂盒，选择位点时参考了中国人群单倍群频率，针对中国人群可获得高分辨率的单倍群结果，单倍群划分细致，推断准确。
(2)本发明采用多重PCR扩增子捕获技术进行引物设计和建库，结合热力学稳定性的算法进行特异性引物设计，采用两步PCR扩增方法来完成目标区域扩增和建库，节省了大量的时间和成本；
(3)本发明可以采用多种样本类型进行文库构建，如口腔拭子、指甲、带毛囊的毛发和精斑等的基因组DNA；由于设计的扩增子片段短，该体系对于降解检材也很友好。
附图说明
图1为本发明一实施例中复合扩增体系所涉及的Y-DNA单倍群基本树(2019年)及选取的Y-SNP数目情况；
图2为本发明一实施例中基于100个样本计算的每个Y-SNP位点的平均测序深度；
图3为本发明一实施例中复合扩增方法的灵敏度测试结果。
具体实施方式
本发明涉及一种基于二代测序技术高分辨率检测Y-SNP单倍群的引物组合物，其包括381个Y-SNP位点的扩增引物；
其中，其中，所述的381个Y-SNP位点均为根据中国人群各主要单倍群所占比例，结合各群体人口数，从Y染色体进化树中选取的下游分支Y-SNP遗传标记，为了保证树的完整结构性，同时加入了许多重要节点主干单倍群，具体包括AF6、P305、P108、M60、M91、M130、F3393、CTS11043、M8、M356、M38、M217、L1373、F3447、F1699、F3918、FGC28881.2、F1756、F3830、Y10420、M48、M86、B90、M504、F3796、F9992、F1067、Z1338、Z1300、CTS2657、CTS11990、CTS8579、M407、F3850、Z12209、F1319、CTS3385、FGC45548、F845、CTS10923、MF2091、P143、M168、M294、CTS3946、JST021355、M174、CTS11577、M15、N1、Z27269、PH4979、PH1991、Y7820、SK541、Z31625、Z31584、P99、P47、M533、Z34359、PH97、Z42599、Z42600、Z42601、Z42602、M116.1、M145、M96、M89、M427、M201、P287、P15、CTS4367、L30、P303、L140、F1329、L901、M69、M52、M82、M578、M170、M429、M522、M304、M267、M172、M410、L26、M47、M67、M9、M526、M20、M22、M27、M317、P326、P256、M231、F3373、CTS3750、CTS11499、F1206、Tat、M178、L708、P298、L392、CTS10760、Z1936、B197、M2019、F1008、M128、B523、P63、B525、F2930、CTS582、M1819、F549、M214、M175、F265、M119、B384、P203.1、F446、F140、CTS3265、F78、F81、F533、F492、F656、SK1568、Z23406、M101、Z23392、CTS8423、CTS52、CTS701、Z23266、CTS716、CTS352、Z23271、M50、F1036、P31、M268、F2320、M1470、PK4、M95、F1252、M88、F2758、F2890、CTS5854、Z23810、Z23781、F789、F838、CTS1456、F993、F417、CTS9996、M176、P49、M122、M324、F113、L127.1、KL1、L467、F1876、F2159、F1867、F852、F2266、L599、Z43961、Z43963、F854、Z43966、F915、IMS-JST002611、F240、F18、F117、CTS5820、F13、F11、F632、F110、F17、F377、F856、F1095、F1418、F1495、CTS7501、F793、Y20951、Y20932、F38、F12、F196、F930、F2685、F1365、F3232、Y15976、Y26383、FGC54486、CTS12877、F2527、F723、F1422、CTS2154、SK1691、PH203、F449、F238、F134、F1273、F724、F1266、CTS498、FGC3750、P201、M188、F2588、CTS445、CTS201、M159、M7、F2680、F1276、CTS6489、F1275、M113、N5、Z25400、F1863、F1134、F1262、Y26403、F1837、F862、F879、F1226、F2859、P164、M134、F450、M117、M133、F42、F438、Y17728、F155、F813、Y20928、F1754、F2137、F1442、F1123、F1369、F310、F402、F1531、Z25853、CTS5492、CTS6987、Z42620、CTS4658、Z25928、SK1730、Z26010、A9462、CTS7634、F317、F3039、Y29861、CTS5488、CTS10738、CTS9678、Z39663、A9457、YP4864、Z44068、F5524、F5525、Z44071、Z44091、CTS4960、F122、F114、F444、F79、F629、F46、FGC16847、F48、F2505、F242、CTS4266、Z26108、F2887、F3607、F3525、CTS3763、A9472、FGC16863、L1360、FGC16889、SK1768、F4249、FGC23868、CTS335、CTS53、CTS6373、A9473、F3386、Y29828、FGC34973、F1739、F743、F748、AM01822、N6、AM01856、N7、F4110、F4068、F4124、AM01845、F717、A16433、F742、F1150、F837、F1025、F1055、F3021、P295、M45、M242、F903、M346、M3、M207、M173、M420、SRY10831.2、M17、Z645、M458、Z91、Z93、F3105、Y7、Z2124、Z2123、S4576、F1345、M343、L754、L389、P297、M269、M479、M124、FGC21706、B254、M184。
下面通过具体实施例和附图对本发明进行详细和具体的介绍，以使更好的理解本发明，但是下述实施例并不限制本发明范围。
实施例中方法如无特殊说明的采用常规方法，使用的试剂如无特殊说明的使用常规市售试剂或按常规方法配制的试剂。
实施例1
本实施例提供一种基于二代测序技术高分辨率检测Y-SNP单倍群的引物组合物和方法，具体的Y-SNP位点筛选、引物设计以及检测方法的步骤如下：
1.Y-SNP位点筛选及引物设计；
根据中国人群各主要单倍群所占比例，结合各群体人口数，从Y染色体进化树中选取不同数量的下游分支Y-SNP遗传标记。为了保证树的完整结构性，同时加入了许多重要节点主干单倍群。参照2019年ISOGG网站(Version:15.58)(https://isogg.org/tree/HaplogroupTr2019.html)，总共筛选出针对中国人群381个Y-SNP位点，构成359个单倍群。将所选的381个Y-SNP位点的人类参考基因组37(GRCh37/hg19)的染色体位置等详细信息上传至引物设计网站(https://design.igenetech.com/)进行多重引物设计。为了获得更好的覆盖度(Coverage)，对引物进行两轮设计和优化。
本实施例所述的扩增引物在Y染色体上的位置及引物序列如表1所示。
表1复合扩增检测体系中扩增子及扩增引物序列
2.建库；
选择多重PCR扩增子捕获技术采用两步PCR扩增方法来完成目标区域扩增和建库。文库构建流程包括：多重PCR反应、磁珠纯化合并产物、接头序列PCR反应、磁珠纯化和文库浓度测量与质检，具体过程如下：
(1)多重PCR：以稀释好的gDNA为模板，使用AI-Mito-Multi Panel试剂盒进行多重片段扩增，扩增反应的总体积为25μL，配置表见表2。
表2.多重PCR反应体系
PCR反应条件：预热105℃；95℃3分钟30秒；14个循环：98℃20秒，60℃3分30秒；72℃5分钟。
(2)利用AMPure XP磁珠纯化PCR产物。
(3)接头序列PCR反应：反应体系见表3。
表3.接头序列PCR反应体系
PCR反应条件：预热105℃；95℃3分钟30秒；9个循环：98℃20秒，58℃1分钟，72℃30秒；72℃5分钟。
(4)进行第二轮磁珠纯化，随后取2μL文库使用3.0 Fluorometer进行浓度测定，记录文库浓度；取1μL文库样本使用Agilent 2100 Bioanalyzer system(HighSensitivity DNA Kit)进行文库片段长度和纯度测量。
(5)将文库混合、变性后加入Illumina MiSeq测序平台进行测序。将测序结果与参考基因组序列比对，采用samtools、GATK等软件寻找SNP和Indel，然后运用ANNOVAR软件对SNP、Indel位点进行注释、确定突变位点对应的基因信息等。
其中，上述复合扩增体系所涉及的Y-DNA单倍群基本树(2019年)及选取的Y-SNP数目情况如图1所示。
实施例2
本实施例为实施例1提供的基于二代测序技术高分辨率检测Y-SNP单倍群的方法的法医学验证工作。
按照DNA分析方法科学工作组(Scientific Working Group for DNA AnalysisMethods,SWGDAM)要求对实施例1构建的方法的测序质量、灵敏度、重复性、准确性、案件检材适用性进行评估并计算法医学参数。
基于100个样本计算的每个Y-SNP位点的平均测序深度如图2所示，文库构建顺利，测序质量较好。
法医学验证的结果显示，该方法的建议DNA投入量至少在1.25ng，但应用于法医实践中DNA浓度极小的微量样本仍有可能获得有意义的单倍群结果，灵敏度测试结果如图3所示；重复性好；特异性好，动物样本和女性样本利用该方法均不能得到有效扩增；准确性高；适用于多种类型检材的DNA分型；在200例汉族人群中共发现7个主干单倍群，其中主流单倍群仍然是O、C、N和D，这与中国人群频率分布一致。细分之下的单倍群可达到93个末端单倍群，HD值高达0.9883。
由上述实施例可知，本发明所述的基于二代测序技术高分辨率检测Y-SNP单倍群的方法可应用于家系排查和对未知样本地理来源的父系族源推断中，为法医学实践提供一种灵敏度高、分辨率高和系统效能强的族源推断技术。
以上对本发明的具体实施例进行了详细描述，但其只作为范例，本发明并不限制于以上描述的具体实施例。对于本领域技术人员而言，任何对本发明进行等同修改和替代也都在本发明的范畴之中。因此，在不脱离本发明精神和范围下所作的均等变换和修改，都应涵盖在本发明的范围内。
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<110> 司法鉴定科学研究院
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cttgttggtc cacagagatg gt 22
<210> 93
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 93
ctttctaaag attggagttg gatctgtg 28
<210> 94
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 94
taccgacaaa gttttaacaa gctatcag 28
<210> 95
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 95
ggaatgtcac ctgagagttg gt 22
<210> 96
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 96
tggacacaag tggctactga ac 22
<210> 97
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 97
caccaggaac tacaaaacaa tgca 24
<210> 98
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 98
aggacaggaa caacttcaaa attctc 26
<210> 99
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 99
attcactctc aggagccagg ta 22
<210> 100
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 100
catctgcttt tgtgtgcctc tg 22
<210> 101
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 101
cttgcatccc ttagaagccc tt 22
<210> 102
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 102
agacacaggg agggtaacat ca 22
<210> 103
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 103
catctgctag agctcagtct aatctg 26
<210> 104
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 104
cagtggctcc caaaaggatg aa 22
<210> 105
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 105
tgactgacct acttggtctc ca 22
<210> 106
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 106
taccgtgccc agccaatttt 20
<210> 107
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 107
ggcacagttc aaacaccagg at 22
<210> 108
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 108
cacatcttga gccattctgg gt 22
<210> 109
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 109
gaagcttctg taaccaggca gt 22
<210> 110
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 110
gcaacaaact gagaccccat ttc 23
<210> 111
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 111
catccatcct tacctgtgtg ca 22
<210> 112
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 112
atggcatcta ttctgctggg tc 22
<210> 113
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 113
tttctgaaca cagccaggga ag 22
<210> 114
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 114
tctgtggttc tgtcccatgt tc 22
<210> 115
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 115
aagtctcaga ctacccaccc tt 22
<210> 116
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 116
gttgagtaca tggtttggct gc 22
<210> 117
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 117
gtgcgaatcg accttatgtt cc 22
<210> 118
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 118
gtgagagttc catgatcccc aa 22
<210> 119
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 119
agtgaacggg gaagagaatg ga 22
<210> 120
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 120
aaggtctggt tgaagggttg tg 22
<210> 121
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 121
gctattgaag cacactggcc ta 22
<210> 122
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 122
cagaagatgg tgaccagcac tt 22
<210> 123
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 123
ttacaggatg gggcaactta cg 22
<210> 124
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 124
ctgactcacc tgcacagtga aa 22
<210> 125
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 125
gctcaatcaa catggggaat gg 22
<210> 126
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 126
accacatctc tcatagtgcc ac 22
<210> 127
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 127
gcacgaggaa agagaagtgt ga 22
<210> 128
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 128
ctgtgcacac aagcatgcat ac 22
<210> 129
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 129
tttcctggag ggtcagaaac ac 22
<210> 130
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 130
atggggtagc aaatgcacgt 20
<210> 131
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 131
taatgaacac cccagggatt gc 22
<210> 132
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 132
tctgaagctc cctgtaatgg ga 22
<210> 133
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 133
agcgtagcca gaaatgtgca 20
<210> 134
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 134
acacaagggc gtaaatgcca 20
<210> 135
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 135
gatgcacgtc ctgaaaagga ga 22
<210> 136
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 136
cttcacaaac ctcgggactg tt 22
<210> 137
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 137
acttgaacct gggatgtaga ggt 23
<210> 138
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 138
tgcaaaactg gaatgtcaac atgg 24
<210> 139
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 139
tctttccttc cctccttccc tt 22
<210> 140
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 140
gagtgctggt ttcactttgt gg 22
<210> 141
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 141
agccttagca aaagagcacg at 22
<210> 142
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 142
gaagttgaca agacagaatt tgttctca 28
<210> 143
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 143
tgaggatgtc cttcgtgtct ct 22
<210> 144
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 144
ttggggagga tcaatgaggc ta 22
<210> 145
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 145
tgaaaccttg tggtcccatc ac 22
<210> 146
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 146
gctatgttgc ccaggctaga tt 22
<210> 147
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 147
gacagacagc aagcactaag ga 22
<210> 148
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 148
tgctagggtt acagacatga gc 22
<210> 149
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 149
tctccaagac acgttctctc ca 22
<210> 150
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 150
ccttgacagg aatagtgaag ggag 24
<210> 151
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 151
caactgcaag tttagtgcct cag 23
<210> 152
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 152
accatccagt ctcatcaagc tg 22
<210> 153
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 153
gaggcaatac cctgatctgc tt 22
<210> 154
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 154
gatggccaaa tggcaacatc tc 22
<210> 155
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 155
agacagagga aagccaaggg ta 22
<210> 156
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 156
acactggact cagactggaa ct 22
<210> 157
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 157
aaggtcctgg aaccagtcct t 21
<210> 158
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 158
aagggtggaa aaggcctcat g 21
<210> 159
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 159
ccacacacct ggacaattgt gt 22
<210> 160
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 160
tctccatctc ctgacctcaa gatc 24
<210> 161
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 161
cacacaggca tagaaaagaa agagtac 27
<210> 162
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 162
cttgcattgt acacactttt tcaagtg 27
<210> 163
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 163
cctggtttct caggaacttg gat 23
<210> 164
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 164
tacgctgtca cctctcagta tct 23
<210> 165
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 165
cagacatgtg acatgtgctg ac 22
<210> 166
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 166
tgtcggacat gtacgacatg tg 22
<210> 167
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 167
agttccatgc ttcctccttg ag 22
<210> 168
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 168
tacccagcag tattagctgt gc 22
<210> 169
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 169
ggagaacaat gagaaagggc agag 24
<210> 170
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 170
aagggcagat catggtctca ga 22
<210> 171
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 171
tccacagtcc ctccataaac ct 22
<210> 172
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 172
tgctgggata aggtacctcc aa 22
<210> 173
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 173
gatttgacac accctggtga ct 22
<210> 174
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 174
aagctccact gagacactca ct 22
<210> 175
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 175
gttgtctgtg ctgagaacag ga 22
<210> 176
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 176
tgactcccat aggacagtga ca 22
<210> 177
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 177
ggaacaggga gggcttatgt g 21
<210> 178
<211> 19
<212> DNA
<213> Artificial Sequence
<400> 178
ttgccctctg aacgggttg 19
<210> 179
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 179
agaacagcag gtgtgagaac tg 22
<210> 180
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 180
cgtttaactc ccgcagtctc at 22
<210> 181
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 181
gcttttcatt gcaggacgct ag 22
<210> 182
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 182
agacacccct ggagcatatc tt 22
<210> 183
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 183
catatggcca aagccaagac ct 22
<210> 184
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 184
gtaccttgtg ttggtgtggg aa 22
<210> 185
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 185
ggcaggttta agggagaaga tcc 23
<210> 186
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 186
ccattccaca tttgcagtgc c 21
<210> 187
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 187
gcagtagcca ctgatacagt gt 22
<210> 188
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 188
atgctgtcta gtcagggagt ca 22
<210> 189
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 189
gtatcgtagc ccaaatcctg ct 22
<210> 190
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 190
cacatgaaca gagcctacca gt 22
<210> 191
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 191
agtccccatt aggacagaac ca 22
<210> 192
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 192
ttgccaggac tatgcttcac ag 22
<210> 193
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 193
cacaagaaag tcccatcgcc ta 22
<210> 194
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 194
tggtgaagtg agtctccctt ct 22
<210> 195
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 195
aagagtcaac tcacctcgat gc 22
<210> 196
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 196
aggtggagtg actgtcctca tt 22
<210> 197
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 197
aggagtgatg tagagtcctg ctt 23
<210> 198
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 198
tggtactgtg ttacgaccta tcct 24
<210> 199
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 199
gcttaagaaa gagaggaggg tcac 24
<210> 200
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 200
gtgactctcc cttttgggtt ctac 24
<210> 201
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 201
ggtctgcagt aagtctcttg aagac 25
<210> 202
<211> 29
<212> DNA
<213> Artificial Sequence
<400> 202
agcagaaaat aagaaggctt tgaagaaag 29
<210> 203
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 203
agtaacatct tccgggtgct tg 22
<210> 204
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 204
tctgagccca tgacctaagt ga 22
<210> 205
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 205
acctgatagc tctctgctga ca 22
<210> 206
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 206
gtctgtctag ggagttaagt ccct 24
<210> 207
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 207
gcaaaaggcc ctcatgatgt g 21
<210> 208
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 208
acagtgtaat ctaaggcccg aag 23
<210> 209
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 209
actggctgta ttcctccact ct 22
<210> 210
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 210
gtgtgtgttg gagaggtgga at 22
<210> 211
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 211
tgtaacctac cacaggtgtg tg 22
<210> 212
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 212
tgctgtccag gtataatggc tc 22
<210> 213
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 213
gcacttgaga aacagctggt tc 22
<210> 214
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 214
ctgtctgcct caccatacaa ca 22
<210> 215
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 215
tgagtcacat tttctgaatg tgaactg 27
<210> 216
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 216
acaggatgca tttcagagat atggtaa 27
<210> 217
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 217
tgagaagagg acactgcatg tc 22
<210> 218
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 218
accacttagt ggtcctctgt ct 22
<210> 219
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 219
ttcatttgtt ctgacgggag ca 22
<210> 220
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 220
gatgtttaat ccacatcaga gatgatgg 28
<210> 221
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 221
tgtctctatc tccttcagtt tcactc 26
<210> 222
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 222
tcagtaactg gtctggttcc tttac 25
<210> 223
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 223
acatcagaag ggacagactc ca 22
<210> 224
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 224
ccaaggcaaa gcaagtgtgt c 21
<210> 225
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 225
tcagggccta tcttgtaggt ga 22
<210> 226
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 226
cctatgtcca ggttccagct tt 22
<210> 227
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 227
gttttacagt cccacagaag tcct 24
<210> 228
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 228
acctgtagtg ctttgactca tctc 24
<210> 229
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 229
ggttcaatgt caccggtcag a 21
<210> 230
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 230
gtgacttcct gattggccct t 21
<210> 231
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 231
tagctggttg ttggtaccag tg 22
<210> 232
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 232
tgactttgac ctgcctctga ag 22
<210> 233
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 233
tcattggtag cttcatgggg at 22
<210> 234
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 234
tggattccct cccattcaca at 22
<210> 235
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 235
gctttgctct gggtagttga ca 22
<210> 236
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 236
gtcagacata tggccagagc at 22
<210> 237
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 237
taagggtcag gaagtctctg ct 22
<210> 238
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 238
ggtagtctgt gttatgctgc ct 22
<210> 239
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 239
gacacagaca tagtcagtga cca 23
<210> 240
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 240
taccctggaa aagtcacacc atc 23
<210> 241
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 241
gacccaaaag gatctgcagg at 22
<210> 242
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 242
aacccatagc taggcagatc ct 22
<210> 243
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 243
gtgtggaatg ggttgatgct tg 22
<210> 244
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 244
cttgtgtgtg gctgtaacca ga 22
<210> 245
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 245
cagcagggaa ttgtgaggtg t 21
<210> 246
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 246
atggtgcagt gctacctcat g 21
<210> 247
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 247
gtgctttcaa ctccatccct ga 22
<210> 248
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 248
gcatccattt ggctccaggt at 22
<210> 249
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 249
ttggtcccct gtctgtatga ct 22
<210> 250
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 250
tggtcttgga tgtggtgtga tc 22
<210> 251
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 251
gcttgcaatc accacagaca tc 22
<210> 252
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 252
caagtcccgt gggtattgga at 22
<210> 253
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 253
tcaaaacgct tcctcctgta gc 22
<210> 254
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 254
cacaaagtag gttctggtgc ca 22
<210> 255
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 255
gcagtttcca gtgatcacct ca 22
<210> 256
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 256
ttgctctcag agatgtcagc ct 22
<210> 257
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 257
gccacttttg cctaacgaca ag 22
<210> 258
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 258
caaggcatga ctaaggcatc ct 22
<210> 259
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 259
tggggtctca ggtatgatgc ta 22
<210> 260
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 260
gaaggacgtg tttcgagact gt 22
<210> 261
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 261
agaaagctca tgcaaggcct ag 22
<210> 262
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 262
gactgtattc cctatagagg ccct 24
<210> 263
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 263
ccagttatgg ccttcttcac ca 22
<210> 264
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 264
gttccttcag gggcctttgt at 22
<210> 265
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 265
tctgtcttct ctgtgggatc ca 22
<210> 266
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 266
aggagagagc atcacctggt ta 22
<210> 267
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 267
gtgtacgcat gcctgtaagt ga 22
<210> 268
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 268
tctcagccag cctctgtatt ca 22
<210> 269
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 269
aaagtctggc ttttgcctgg ta 22
<210> 270
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 270
catgtaaagt gcaaaccgcc tt 22
<210> 271
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 271
ccttcacatc atgaagggga tctc 24
<210> 272
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 272
gagtacttcc tcatgaacac cgag 24
<210> 273
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 273
atgtttctgg gactccagga ca 22
<210> 274
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 274
cttgtctctg acgtgtgcag aa 22
<210> 275
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 275
tgcaccttct gtgtcatctg tc 22
<210> 276
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 276
agctgagttg gtgcttcaga tc 22
<210> 277
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 277
acagaggcag gctagaaatc tg 22
<210> 278
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 278
gagaaggtca cttccgttga gt 22
<210> 279
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 279
caatgtgggg atggagttgg aa 22
<210> 280
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 280
tacccctcca ttgtcttggg at 22
<210> 281
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 281
ccttctgctg gagtcaaagc aa 22
<210> 282
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 282
tgactgcctc tcaccacaga tt 22
<210> 283
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 283
gaagcactgc agtgcaatag c 21
<210> 284
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 284
gttctcttcc aagtggggct t 21
<210> 285
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 285
agacactgtg gattcctcct ga 22
<210> 286
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 286
gactgcagtg tgccacagat ta 22
<210> 287
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 287
caggcaaaag ccagacactc 20
<210> 288
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 288
ctcatgggaa tcccttgcgt 20
<210> 289
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 289
tgaacccatc ctctgcttca tg 22
<210> 290
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 290
tccatggagg acagatcact ga 22
<210> 291
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 291
ttgcaaactt ccagtccctc ag 22
<210> 292
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 292
aaggagatgc ccaaggatct ca 22
<210> 293
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 293
attcacagag cccagctctg a 21
<210> 294
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 294
tggtgatgga tgacatcatg gc 22
<210> 295
<211> 34
<212> DNA
<213> Artificial Sequence
<400> 295
agagaatatt tcatgaaact acattataca gtac 34
<210> 296
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 296
gaaccattgt gaggagaaag ttatactt 28
<210> 297
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 297
cttttgcatg tgtggatatg tgtct 25
<210> 298
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 298
cagtataggc caacactggc tta 23
<210> 299
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 299
cttgaaatgc ctcctccacc a 21
<210> 300
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 300
tcacaatcag gcctgatgtc c 21
<210> 301
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 301
ccccactggg attggaatat actc 24
<210> 302
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 302
agtccaaagt gagagaacac gg 22
<210> 303
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 303
agtgtactga gtggtgacac cta 23
<210> 304
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 304
gacttgtgct tggtcagtca aac 23
<210> 305
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 305
atcccttgca gaatcccttt cc 22
<210> 306
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 306
tagggcagga acaattgact gg 22
<210> 307
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 307
caaaaaagag cccacattgc caa 23
<210> 308
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 308
tgtggcataa ttttgagggc tct 23
<210> 309
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 309
agcagcattt tatcctccaa gtca 24
<210> 310
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 310
tggaagacaa tggatgtcaa gtgt 24
<210> 311
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 311
agagacacta gccgtcttgt ct 22
<210> 312
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 312
ggaactcggg gaagaagttt gt 22
<210> 313
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 313
tgtttgatgg ttcagcttcc aga 23
<210> 314
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 314
tctcctttct gtcagcttta gctt 24
<210> 315
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 315
ggcagaggtg gaagaaaatt cac 23
<210> 316
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 316
tcaccacttg caaggatact cag 23
<210> 317
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 317
ggaagaggcc aaacaaacat cg 22
<210> 318
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 318
tgactggaag tgtgtcttgg ac 22
<210> 319
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 319
agcaaaaccc cagatgacag ag 22
<210> 320
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 320
agtgacatct gacctctcac ca 22
<210> 321
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 321
ttagtcccta ggggacactt gg 22
<210> 322
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 322
gacagggtta cttgacctca gc 22
<210> 323
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 323
gactcttgcc caaaactggg at 22
<210> 324
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 324
ttcctgcaag tgtgtctgga tc 22
<210> 325
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 325
ctgaatgtgg gacaagatga gga 23
<210> 326
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 326
gatgtgatcc tgctatattg cgc 23
<210> 327
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 327
gggaaggacc gtaaatctgc tt 22
<210> 328
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 328
cagtgggaaa ggagcatcct ta 22
<210> 329
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 329
cattccattg catcccctac ct 22
<210> 330
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 330
cagagccaga ttccaaaatg cc 22
<210> 331
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 331
aaccctgagg agacatagcc at 22
<210> 332
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 332
gtcatccaag gagcaacaca gt 22
<210> 333
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 333
agatgtcagg accaactctc ca 22
<210> 334
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 334
tctccaatca agtccctccc a 21
<210> 335
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 335
gtggtgattc ctcaaagatc tgga 24
<210> 336
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 336
catgtccctg caaaggacaa ttc 23
<210> 337
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 337
aacacttact gagcccctgc ta 22
<210> 338
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 338
cacacatggc agaaggaatg ga 22
<210> 339
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 339
tcttctccaa gagcctgaag ga 22
<210> 340
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 340
tgtgcacatg tgcccttgaa 20
<210> 341
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 341
tgtgcccttg ccatttttgc 20
<210> 342
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 342
ctgagtatca cgtgagagta gcc 23
<210> 343
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 343
tacatcccag tccagtcacc tt 22
<210> 344
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 344
aaggcttaga gaggggagca a 21
<210> 345
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 345
tctcctttac tcaggcctga ca 22
<210> 346
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 346
ttggcctcat tcccttctca ac 22
<210> 347
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 347
tcagtccata ctagtgggcc at 22
<210> 348
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 348
cttgtaagga agccatccca ca 22
<210> 349
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 349
tttggggaac aggtaggtgg ta 22
<210> 350
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 350
agctacttgg gaggctgata ca 22
<210> 351
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 351
atagacgaga ggagggggag a 21
<210> 352
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 352
catccatcca tgaactgctt ctact 25
<210> 353
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 353
aatgaggccc accacattat gg 22
<210> 354
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 354
cacagccaaa taccagtcgt ca 22
<210> 355
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 355
gaggagaaag cagagctcat acac 24
<210> 356
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 356
gcattcacgc atacatgcag tc 22
<210> 357
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 357
gccaagagat gcatagcaag gt 22
<210> 358
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 358
catggagtct tggcagcact at 22
<210> 359
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 359
cccagaaagt tgagacagct ga 22
<210> 360
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 360
ccattttttc tgaagcccgc tac 23
<210> 361
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 361
ttaggcatgt ggacactcac ag 22
<210> 362
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 362
agcatgccta cagcagacta ag 22
<210> 363
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 363
ggtacaggtt aatgcagggc tt 22
<210> 364
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 364
ccaagtaatt ctttgttgtt cagcagt 27
<210> 365
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 365
gctgtgtgga gtatgtgttg ga 22
<210> 366
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 366
gacaaatatc ctgccccagc tt 22
<210> 367
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 367
atccaggtca tgcagtttgc t 21
<210> 368
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 368
aagcgagcac agatgtaagc a 21
<210> 369
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 369
gggttccaga ttttgcatga tcg 23
<210> 370
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 370
gtgagacaca acccacactg aa 22
<210> 371
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 371
gatctgctag caaagttggc ttg 23
<210> 372
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 372
tctgattgga agaaacaaca atgacc 26
<210> 373
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 373
ccaacagagt gagagtctgt ctg 23
<210> 374
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 374
tagcactctt ccacccactc t 21
<210> 375
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 375
aggccaggag aatgatgtga ac 22
<210> 376
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 376
tgtccatcgt tcatgtcagc at 22
<210> 377
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 377
tcctttgcag atagaacagc tgtag 25
<210> 378
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 378
tgcaaaggca aaaaccaacc tc 22
<210> 379
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 379
tggactctga gtgtagactt gtga 24
<210> 380
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 380
gacaagaaga acttcagcat tggg 24
<210> 381
<211> 29
<212> DNA
<213> Artificial Sequence
<400> 381
cagctgtctt attaaaagga ggtttgtag 29
<210> 382
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 382
ccaagaagca ccaaagtgaa agaaa 25
<210> 383
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 383
ctttcccgca tccaaagttt acc 23
<210> 384
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 384
ctcactagta attgctgtgc cca 23
<210> 385
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 385
ggtggcttat gttgttggtg tc 22
<210> 386
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 386
gtagcaccag catttttgag tcc 23
<210> 387
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 387
gtatgatgca ctgtgcgttc ttc 23
<210> 388
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 388
gaaggacaag acccatcttg ca 22
<210> 389
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 389
gcagttgtcc tgttgcttac ca 22
<210> 390
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 390
gttcatctcc tagccattcc actg 24
<210> 391
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 391
ttaggagcca gatgaccagg at 22
<210> 392
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 392
cactccatgt tggtttggaa cag 23
<210> 393
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 393
aggacattgt tcacccacct tg 22
<210> 394
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 394
tgaggaagtg ctcagaagag gt 22
<210> 395
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 395
ttgtgcaaaa aggtgaccaa gg 22
<210> 396
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 396
tccaatctca attcatgcct cct 23
<210> 397
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 397
gcacagtact cactttaggt ttgc 24
<210> 398
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 398
agcttctcct ggaccatctg tata 24
<210> 399
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 399
caacatgggt aattcggttg ttacc 25
<210> 400
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 400
tcagcttcat ccaacactaa gtacc 25
<210> 401
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 401
ttggcagtgc ttcaaagagg t 21
<210> 402
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 402
agtggaagct ccttgagact ct 22
<210> 403
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 403
ggagctaaac aacaggtcca ca 22
<210> 404
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 404
actgcataat ggtagggatt gagc 24
<210> 405
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 405
ggatgggctg aaacaaaggg at 22
<210> 406
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 406
aggtcgtggg ttttctggaa tg 22
<210> 407
<211> 31
<212> DNA
<213> Artificial Sequence
<400> 407
ctaaaaatac aaaaaattag ctggaagtgg t 31
<210> 408
<211> 31
<212> DNA
<213> Artificial Sequence
<400> 408
tcgtttttgt tcattttctt tcttgagata g 31
<210> 409
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 409
ccaaagaaag gccagtatct cca 23
<210> 410
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 410
agctgctgtg gctttcatca 20
<210> 411
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 411
ggtagaaact ggaaggtgcc tt 22
<210> 412
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 412
cgttgtgttt gacaccacag aa 22
<210> 413
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 413
catgcccatg gtccaattgt ac 22
<210> 414
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 414
ccatggaaat gccacttcac tc 22
<210> 415
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 415
actctctgaa tcaggcacat gc 22
<210> 416
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 416
ctcttgcagc attttcagtt agcc 24
<210> 417
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 417
ggggcaaatg taagtcaagc aag 23
<210> 418
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 418
cttccaggaa ggacatgcct tt 22
<210> 419
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 419
cctgaaactg gctggaaggt ag 22
<210> 420
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 420
ctacttggaa ggctgacatg gg 22
<210> 421
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 421
gcttccacaa gatgcagagt ca 22
<210> 422
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 422
aggtcccaga cttgatgaag gt 22
<210> 423
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 423
gtttgggcca gtatcttcaa ctg 23
<210> 424
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 424
aggagagatc aaagttctgc cc 22
<210> 425
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 425
tccttctcct acaaaggcca ga 22
<210> 426
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 426
gtgagccaag aaggaggaat ca 22
<210> 427
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 427
tgcatcctgt ctaagccatc ac 22
<210> 428
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 428
agcagattgt gatgcacata atgtatg 27
<210> 429
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 429
ctcagaggct caaccattgg aa 22
<210> 430
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 430
ccttaagtct gggtgggata aagg 24
<210> 431
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 431
aggttggtcc tgggaaaaag tc 22
<210> 432
<211> 29
<212> DNA
<213> Artificial Sequence
<400> 432
tgcagaaaaa catgaccaga atttttatc 29
<210> 433
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 433
gtctcatgac acatgacccc tt 22
<210> 434
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 434
ctgacaaatc acctctggtc ca 22
<210> 435
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 435
tttcccccca tcggatttta cc 22
<210> 436
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 436
aaggggagag ggagcattag g 21
<210> 437
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 437
cttgccctca caatttcact catc 24
<210> 438
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 438
ctcttgaact ggctttcagg aaac 24
<210> 439
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 439
tagcacgtat tctgccaggt ct 22
<210> 440
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 440
tctcccttgg aaaacctctg ga 22
<210> 441
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 441
aagtgtgtag ggccatgtga ga 22
<210> 442
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 442
gtgtctttgc gtgtgtgtgt tc 22
<210> 443
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 443
gttgggcaac aactgttcta gc 22
<210> 444
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 444
cctatctgtg gagcctgcat tt 22
<210> 445
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 445
gtcaggagtt cacaacacac ct 22
<210> 446
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 446
cagtgatgca atcttgggtc ac 22
<210> 447
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 447
tgacccatag ttaccccaga ca 22
<210> 448
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 448
ttggaggtgg gattccctga ta 22
<210> 449
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 449
cactttggga tgcagaatca tatgg 25
<210> 450
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 450
taggtggaat gtcctctgtt gcc 23
<210> 451
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 451
ctgtgaccca aacacctacc aa 22
<210> 452
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 452
ggttcaccaa ctgatgaaga tgc 23
<210> 453
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 453
ggaagcagct ttgaagacaa gc 22
<210> 454
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 454
agagatctgg cccaagacag aa 22
<210> 455
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 455
gggtctcagc agtgttttac ct 22
<210> 456
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 456
taggtgttgg gttgggtttg g 21
<210> 457
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 457
ttgccttttt tccttgccca ag 22
<210> 458
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 458
aggaaaggca agggtgttga tt 22
<210> 459
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 459
ctcccaaagt tagcttggcc ta 22
<210> 460
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 460
gagtgcctgt ttcaggtaca ga 22
<210> 461
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 461
gggcaagtct tctttcttgg gat 23
<210> 462
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 462
gacaaaagga gaaatgccac agc 23
<210> 463
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 463
tctaagtgta gtgtccagcc ctac 24
<210> 464
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 464
ctcacctcct tatcctatcc cctt 24
<210> 465
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 465
tcctcctctg aatgtgtgtc ct 22
<210> 466
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 466
agagtgagag gaaggccata ca 22
<210> 467
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 467
gaggactttg aatgttccca acc 23
<210> 468
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 468
gtgtgcctta caaccatctc aac 23
<210> 469
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 469
ctgtttccat cgatggtagt ttgg 24
<210> 470
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 470
caactgcaac catcacgaca tag 23
<210> 471
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 471
ctcaggaagc tgtatctgct tacc 24
<210> 472
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 472
accaccccaa tgatccaatc ac 22
<210> 473
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 473
tggaggcctc ataatgtcct ca 22
<210> 474
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 474
acacttgtcc aagaagtggg atc 23
<210> 475
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 475
ctcaggtatg cttgtgtcca gt 22
<210> 476
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 476
gttgtccact acagctgttt cg 22
<210> 477
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 477
gaatgagagc aggcataatc tggt 24
<210> 478
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 478
ttcccctccc tcattttcta tcca 24
<210> 479
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 479
caatcatggc tccctgaagc at 22
<210> 480
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 480
ctctgttgtt tctgggcatg gt 22
<210> 481
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 481
gaatccctga tttctcattt actgctag 28
<210> 482
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 482
gaactcctga gctcaagtga taca 24
<210> 483
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 483
tccttcttcc atcctcctgg tt 22
<210> 484
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 484
tctgctagtg cactgcttca ag 22
<210> 485
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 485
ccctttggtg tagtgcatca ac 22
<210> 486
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 486
tctcagcacc ttagctaatg agag 24
<210> 487
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 487
aatcccaggc atcaacctct ga 22
<210> 488
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 488
cttcaaagca cccacatcct gt 22
<210> 489
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 489
accccttgga aaggaaaaga cag 23
<210> 490
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 490
gacacatatg gtttagaggt gcct 24
<210> 491
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 491
atgtgagcgt ctctccattc tg 22
<210> 492
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 492
atgcttccat gggtagaatg gg 22
<210> 493
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 493
acaaggtgga gaaggagatg act 23
<210> 494
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 494
tccacctctc agattcagta cagag 25
<210> 495
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 495
tcttggagtc cttgacctag gtt 23
<210> 496
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 496
gtctgcctta tttcttaagg tctgc 25
<210> 497
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 497
gcactcctca agatgagaaa tgttg 25
<210> 498
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 498
gaactgagag gttgcatttc acatac 26
<210> 499
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 499
ggcttgctgc atccattgac 20
<210> 500
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 500
acacagggag gggatcatca 20
<210> 501
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 501
ttcaaaaggg gaggtggtgt ac 22
<210> 502
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 502
gttggaaact ccaccctggt aa 22
<210> 503
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 503
catagcaggg ctgttttcaa gc 22
<210> 504
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 504
gagagagaag gttggtgttg ca 22
<210> 505
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 505
tgactgtgac atctgcctcc ta 22
<210> 506
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 506
gatgcacaga gacacggtga a 21
<210> 507
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 507
agaggaaggc cactgattag ca 22
<210> 508
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 508
tctgctactc acacaagagg tgt 23
<210> 509
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 509
gcaacatggg attggttcac tc 22
<210> 510
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 510
cttcccactc tgctggtctt aaa 23
<210> 511
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 511
ggtatacacc agtcctaact gagga 25
<210> 512
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 512
gtctcacagt tggctacatg tca 23
<210> 513
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 513
actgagattc tcccaagttt gct 23
<210> 514
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 514
tgtgaaacag tgagattccg tct 23
<210> 515
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 515
gtcctaccat tccagatgga acc 23
<210> 516
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 516
tacgtgagcc cccaaaatct g 21
<210> 517
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 517
acaggtgtct gcactcaatc tc 22
<210> 518
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 518
caacaaaggc agggaaaacc aa 22
<210> 519
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 519
ggggcaaggg aagaaaaaga ca 22
<210> 520
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 520
tccacagctc agcttctcat ct 22
<210> 521
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 521
ccaacctctc acattttgtc catg 24
<210> 522
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 522
ggatgtttgg gaacagtccc ata 23
<210> 523
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 523
cagcaggtgg attgctggat 20
<210> 524
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 524
gttctatgac cgtggtggtc tc 22
<210> 525
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 525
acccacatcc aaggtcattt cc 22
<210> 526
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 526
gacaggctac tacagcactt acag 24
<210> 527
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 527
ctcttggctc tcattctctc ttgtc 25
<210> 528
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 528
gcaatgacaa atacacttgc ctagc 25
<210> 529
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 529
tcatctaccc ttgtggctca ca 22
<210> 530
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 530
tgagtgattt tctgctgctg gg 22
<210> 531
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 531
ccctctaaaa ctgcaagagg tgt 23
<210> 532
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 532
tagggcccaa tctctcaact gta 23
<210> 533
<211> 31
<212> DNA
<213> Artificial Sequence
<400> 533
actaaaaata aagaaaatta gctgggtgtg a 31
<210> 534
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 534
catcactcac agtgttgctt atctttt 27
<210> 535
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 535
cccagatatt acatgggaca tgct 24
<210> 536
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 536
caaggtacat tcgcgggata aatg 24
<210> 537
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 537
tctacacatc tcaggctggc ta 22
<210> 538
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 538
cgatgtgggg gaaacatcaa ac 22
<210> 539
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 539
ccacgctttc tgttccttct ct 22
<210> 540
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 540
agtgtgtcca ggacagaaga ga 22
<210> 541
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 541
aagggaaatc tgagtcccca ca 22
<210> 542
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 542
gacacggatg catatgtgtg gt 22
<210> 543
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 543
tcctggatat gcaagagggg aa 22
<210> 544
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 544
ctgtgcaaag ggaggaactt ca 22
<210> 545
<211> 29
<212> DNA
<213> Artificial Sequence
<400> 545
ggaaacatat caagattctt ccattaggt 29
<210> 546
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 546
acacgtacaa cagctgcatt tg 22
<210> 547
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 547
aaggatcctg ctgctttcct ac 22
<210> 548
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 548
cttgattagg agccagagtg ct 22
<210> 549
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 549
cacaagtgca catttgggga tg 22
<210> 550
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 550
ttggtgctca gttccatgca 20
<210> 551
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 551
catgttcctt ccttcacaca tgc 23
<210> 552
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 552
gcaatggtgg atccatgtca cta 23
<210> 553
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 553
ggaagttgat gtatggggtg aga 23
<210> 554
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 554
ctgtccgaaa ttctccacaa agg 23
<210> 555
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 555
tccaatctgt cctcacagga ct 22
<210> 556
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 556
catcaaggcc ttgggtagga tt 22
<210> 557
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 557
gttcctcaca caagctcacc a 21
<210> 558
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 558
cactcagcat cactggaatg ga 22
<210> 559
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 559
ttcacagtct cagcgatgtg ac 22
<210> 560
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 560
tgacctggat accacaaagc ac 22
<210> 561
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 561
tcctaaactc gtccctcagt ca 22
<210> 562
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 562
gaactcaggc ttaactctgc ct 22
<210> 563
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 563
tcttgtcttt ttacccagct tacca 25
<210> 564
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 564
actgaagtac acagacaaat gacact 26
<210> 565
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 565
acctcttctg tgggaagttt gag 23
<210> 566
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 566
acagcagcaa tgtggctttt c 21
<210> 567
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 567
cagctggggt tcagttttag act 23
<210> 568
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 568
taccaagatg ccctgggatt tg 22
<210> 569
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 569
gggttgggga ctggtaatat agag 24
<210> 570
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 570
ctgatccatc atcctgggta gtac 24
<210> 571
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 571
tcagtaagtt gggaagcagc aa 22
<210> 572
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 572
ctgtctaagc tacttgctgg ca 22
<210> 573
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 573
gttcctccca ctcctttttg gat 23
<210> 574
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 574
caactttctc caaagcctag ggt 23
<210> 575
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 575
ccagttcctg caaagttagg aga 23
<210> 576
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 576
taggtagata cgaggtctcg ctc 23
<210> 577
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 577
gcacagagca caaatcctga ac 22
<210> 578
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 578
gttgactctg tccctgcttc at 22
<210> 579
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 579
tcccttttct tctggggttt gg 22
<210> 580
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 580
tgcataatga agtaagcgct acct 24
<210> 581
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 581
cagagtttgt ggttgctggt tg 22
<210> 582
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 582
gctaagcact gaagacaccc aa 22
<210> 583
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 583
gtacagttgg ccctttgtgt ct 22
<210> 584
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 584
aacctacaca catcctcccg ta 22
<210> 585
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 585
agcctcctaa agtgtgggga t 21
<210> 586
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 586
gatcatcttt gggcagccac t 21
<210> 587
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 587
cagagactcc gaaagtctgc at 22
<210> 588
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 588
ggaagcgtca gaagcgttaa ag 22
<210> 589
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 589
ctggcactag cagtttgtac ca 22
<210> 590
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 590
actctccaca aaacagctgg ag 22
<210> 591
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 591
cccttccact cttagcttga ca 22
<210> 592
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 592
gtcttccagg gtaacagaaa atgg 24
<210> 593
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 593
catccctggc caataaaggt gt 22
<210> 594
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 594
agcagtgctc aagattcatc agt 23
<210> 595
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 595
tgttatgggt tattccaatt cagcat 26
<210> 596
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 596
tagggaaatg ccaaggtaaa tgact 25
<210> 597
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 597
gttgaggttg cacaaactca gt 22
<210> 598
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 598
acacagagca agtgactctc aa 22
<210> 599
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 599
agatagcttt attcagattt tcccctga 28
<210> 600
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 600
catgggtttt tcttcctttc taccatt 27
<210> 601
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 601
tcacagttca attcttgtct gttgc 25
<210> 602
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 602
gtcttgaaaa gcaaaagaga actgc 25
<210> 603
<211> 33
<212> DNA
<213> Artificial Sequence
<400> 603
tgtgaatctt attttatcta ctcaacttcg ttc 33
<210> 604
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 604
ccatgctaac agagatctga ctgaat 26
<210> 605
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 605
tttcccaggg ctactgtttt cc 22
<210> 606
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 606
gaactgttcc ttctggcctg tt 22
<210> 607
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 607
gactccaggc tgtttcctat acag 24
<210> 608
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 608
tctgaagaag ggcaacagaa cc 22
<210> 609
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 609
gtgctagtgg ggtttgaaca ac 22
<210> 610
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 610
cctttgtttt ttcctcccag tcc 23
<210> 611
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 611
gggtgtggac tttacgaacc aa 22
<210> 612
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 612
caaggattca ggctcttcag ct 22
<210> 613
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 613
aggccactct tggtcttatc ct 22
<210> 614
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 614
taggacctgg ttcatcagct ca 22
<210> 615
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 615
gggtgttcaa cccacatata ctgt 24
<210> 616
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 616
cgttacagag acacagacca aca 23
<210> 617
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 617
tgtaggaaga gtggaaaggc tct 23
<210> 618
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 618
gagtagaggg acaaagttgt ggag 24
<210> 619
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 619
gagaattttt ggtcaaataa aaggccat 28
<210> 620
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 620
acgtctaacc acaatgtagt tgaaaatt 28
<210> 621
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 621
cagaaaacaa aaacaacagc aacga 25
<210> 622
<211> 29
<212> DNA
<213> Artificial Sequence
<400> 622
cctttttctg ctcatacttc ttttatcct 29
<210> 623
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 623
gcttcaggag gctgtttaca ct 22
<210> 624
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 624
gacttgactg aaccagaggc aa 22
<210> 625
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 625
acctccactt tgctcatcct tc 22
<210> 626
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 626
cttgggcctt atggctaagg at 22
<210> 627
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 627
tataggctat ggcctaggtg ct 22
<210> 628
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 628
tggtcttcca tgtgatggtc ag 22
<210> 629
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 629
gccacatcac tcccattatt tgc 23
<210> 630
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 630
catctgactg ctgtagggac aaa 23
<210> 631
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 631
ccgttaagca aaaatccccc tac 23
<210> 632
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 632
aacggaaatg ccaagaatcg tac 23
<210> 633
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 633
ctggaattcc aagctggtga gt 22
<210> 634
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 634
agtcttggca gaatagctgc tc 22
<210> 635
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 635
ccagcaatag tgttgcacct tc 22
<210> 636
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 636
ctgtttaaag cactctggcc ac 22
<210> 637
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 637
tgatttcggg gtactgactc tga 23
<210> 638
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 638
cacttggact gagttgatgt cca 23
<210> 639
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 639
gaataagaga ggagagccac cttc 24
<210> 640
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 640
ctgtgagcac atgagagagc at 22
<210> 641
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 641
agtgagccac catcttcaag ac 22
<210> 642
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 642
agagggtggt tcctatggtc tt 22
<210> 643
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 643
cgcctgtagt tccagttact caag 24
<210> 644
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 644
actccatgtg agaaacttcc ttgtg 25
<210> 645
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 645
gctcttaaag cacagggagg tt 22
<210> 646
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 646
acagcaaggc tgattgctaa gt 22
<210> 647
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 647
atgtgagaag cacctcttcc ca 22
<210> 648
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 648
gctcctcaca tccccgatga t 21
<210> 649
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 649
gggacactac acagttcatt ctgt 24
<210> 650
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 650
gcttaaccga atgtctcaaa tggac 25
<210> 651
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 651
tcatgcctag cctcattcct ct 22
<210> 652
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 652
tagcctggat ggtcacgatc t 21
<210> 653
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 653
ctggatagcg gattcgatgg aa 22
<210> 654
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 654
atcctccaga gaacttggga ct 22
<210> 655
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 655
gcttaaaaga gattgatcgg tgca 24
<210> 656
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 656
gactacaggt gtatgctggc taat 24
<210> 657
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 657
tgtagaccat ctgtcctcat cca 23
<210> 658
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 658
ggtaagcaaa caattgtgct cca 23
<210> 659
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 659
tggggtaggc aaagaaaagc a 21
<210> 660
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 660
tcaggcttct agcttcatct gc 22
<210> 661
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 661
ggctacatgg aaatggttgg cta 23
<210> 662
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 662
cttgaacttg gcaggcgtag aa 22
<210> 663
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 663
gcctgataat catgctgggt tc 22
<210> 664
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 664
aaccgaggca caaaggatga 20
<210> 665
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 665
aatgccttca ctcagcactc ca 22
<210> 666
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 666
aggtgttggc ttttaagcag gc 22
<210> 667
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 667
cacagctgtt ttgagttctc catc 24
<210> 668
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 668
gaaggatcgc taataagccc agat 24
<210> 669
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 669
actcagtggc aactgtgaag tg 22
<210> 670
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 670
ctaggccccc tattggtgag tata 24
<210> 671
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 671
gcagctgggt ttgatactca ga 22
<210> 672
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 672
gtctctactt tctgggcagg aag 23
<210> 673
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 673
cctgttaggt gattgccaga ga 22
<210> 674
<211> 19
<212> DNA
<213> Artificial Sequence
<400> 674
ttgctggtgc acaaacgtg 19
<210> 675
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 675
gcttcccttt tctctgtgct tc 22
<210> 676
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 676
cctaggtaag tgtgtgctcc at 22
<210> 677
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 677
gagagattag gttgcctgcc at 22
<210> 678
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 678
atccagcagg gtatgtgatg tc 22
<210> 679
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 679
cagcaggtag gctttgaagt gt 22
<210> 680
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 680
atcctgaacc tactcgggga tt 22
<210> 681
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 681
gggcatccga cttactattg ct 22
<210> 682
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 682
tccttgaggg tcagaggaag aa 22
<210> 683
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 683
gaaacatatc ggaatttctt ctcaacct 28
<210> 684
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 684
ttccctagga gctcatacaa ctatct 26
<210> 685
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 685
tgtctaccat ggggcttttc tg 22
<210> 686
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 686
gaagcctagg atgcccttga aa 22
<210> 687
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 687
gcaaacccag caatcagtct tg 22
<210> 688
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 688
actgttgcac cctctgagta ga 22
<210> 689
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 689
tctttgctat ctgctgggtg tg 22
<210> 690
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 690
tctgctcata cgcatgttga gg 22
<210> 691
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 691
gcacactcaa cagacagtga ag 22
<210> 692
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 692
tgtgtatgag agcctcaatc ca 22
<210> 693
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 693
gaggaaaagt ggccattgat gc 22
<210> 694
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 694
gcttgctcat ggggccatat at 22
<210> 695
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 695
actaaaaata tgtcagcctc tgtaagg 27
<210> 696
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 696
tccaatatgg gaccatattt cccaaat 27
<210> 697
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 697
cattccaatt cacaatgtga attgtgtc 28
<210> 698
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 698
tggtaaaaaa gtgttagact aaggccta 28
<210> 699
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 699
tacttagagg cagggtgttg ct 22
<210> 700
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 700
atggcagttt tgcaggggtt 20
<210> 701
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 701
cttgcaagct tttccaaaat tgct 24
<210> 702
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 702
gcattgatta atgccattta gacgc 25
<210> 703
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 703
gatgtggctg tcttgactga ga 22
<210> 704
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 704
gtctagggga ctggtgatta ttgg 24
<210> 705
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 705
ggagtgcaaa ttgacaccat cc 22
<210> 706
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 706
ctcatgcagt ggattccact ga 22
<210> 707
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 707
ttgtcatggt tccagccttt cc 22
<210> 708
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 708
gcttttctca ctccatggct ga 22
<210> 709
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 709
cactgcttag tgagtgtccc tt 22
<210> 710
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 710
ctgtaaaact cactgcccag gt 22
<210> 711
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 711
cccttaatgg aaggcatgtg ct 22
<210> 712
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 712
agacggtttg ttgacttgtc ctac 24
<210> 713
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 713
gccagagtca gaaaagagtg cta 23
<210> 714
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 714
tcctgacttc tggagtgaaa acc 23
<210> 715
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 715
ggatgtgtgc attggtctag caa 23
<210> 716
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 716
ggtttactag cacagagctg caa 23
<210> 717
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 717
actagagacg gggttttgca 20
<210> 718
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 718
accttaccct gataccttcc ag 22
<210> 719
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 719
gaagacaaaa gggcaagcga ag 22
<210> 720
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 720
tggaattggg gtgtgttacc tg 22
<210> 721
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 721
tcagtatatg gatggggcag ca 22
<210> 722
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 722
cttcataacc accctgtgca gtac 24
<210> 723
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 723
gccagtctca ctcaattgta cac 23
<210> 724
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 724
ttccctttaa gaagccaaga agc 23
<210> 725
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 725
ccagcttcaa aaacagcaaa gga 23
<210> 726
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 726
tggatgcaga cagaagttga cat 23
<210> 727
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 727
accactgtgg agagatactt ttgatc 26
<210> 728
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 728
gttcaaaata catgagactg cctacc 26
<210> 729
<211> 34
<212> DNA
<213> Artificial Sequence
<400> 729
tcttccctta atagttggat atgattttat gttc 34
<210> 730
<211> 31
<212> DNA
<213> Artificial Sequence
<400> 730
agcaaaactt ttcctataga agcaaagata a 31
<210> 731
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 731
tgccaagatg tcagcttgtg a 21
<210> 732
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 732
cagagtgaga ctcttgtctc aaaac 25
<210> 733
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 733
ttaggccctg ttcttgtgag ag 22
<210> 734
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 734
ctctcaaagc cagtcatggc aa 22
<210> 735
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 735
gttcacctgt gcaagatcat gttg 24
<210> 736
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 736
taggcctcca cttcataatc tgga 24
<210> 737
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 737
tcagttttca gttcactgtg aaaatctc 28
<210> 738
<211> 30
<212> DNA
<213> Artificial Sequence
<400> 738
gcattaggag ataaatctaa cgtaagtgac 30
<210> 739
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 739
ctggccattg tgaactcaag tc 22
<210> 740
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 740
taggtgttag ctcttccaca gc 22
<210> 741
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 741
tgtacagctg gaggaagtac ct 22
<210> 742
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 742
agcttctcct ctggagtcga aa 22
<210> 743
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 743
agctggagga agtacctgct a 21
<210> 744
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 744
agcttctcct ctggagtcga 20
<210> 745
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 745
gaagaaattt gtacagctgg aggaag 26
<210> 746
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 746
gcatatttct tacagagagc tgcc 24
Claims (9)
1.一种基于二代测序技术检测Y-SNP单倍群的引物组合物，其特征在于，包括381个YSNP位点的扩增引物；所述381个Y-SNP位点包括AF6、P305、P108、M60、M91、M130、F3393、CTS11043、M8、M356、M38、M217、L1373、F3447、F1699、F3918、FGC28881.2、F1756、F3830、Y10420、M48、M86、B90、M504、F3796、F9992、F1067、Z1338、Z1300、CTS2657、CTS11990、CTS8579、M407、F3850、Z12209、F1319、CTS3385、FGC45548、F845、CTS10923、MF2091、P143、M168、M294、CTS3946、JST021355、M174、CTS11577、M15、N1、Z27269、PH4979、PH1991、Y7820、SK541、Z31625、Z31584、P99、P47、M533、Z34359、PH97、Z42599、Z42600、Z42601、Z42602、M116.1、M145、M96、
M89、M427、M201、P287、P15、CTS4367、L30、P303、L140、F1329、L901、M69、M52、M82、M578、M170、M429、M522、M304、M267、M172、M410、L26、M47、M67、M9、M526、M20、M22、M27、M317、P326、P256、M231、F3373、CTS3750、CTS11499、F1206、Tat、M178、L708、P298、L392、CTS10760、
Z1936、B197、M2019、F1008、M128、B523、P63、B525、F2930、CTS582、M1819、F549、M214、M175、F265、M119、B384、P203.1、F446、F140、CTS3265、F78、F81、F533、F492、F656、SK1568、Z23406、M101、Z23392、CTS8423、CTS52、CTS701、Z23266、CTS716、CTS352、Z23271、M50、F1036、P31、
M268、F2320、M1470、PK4、M95、F1252、M88、F2758、F2890、CTS5854、Z23810、Z23781、F789、F838、CTS1456、F993、F417、CTS9996、M176、P49、M122、M324、F113、L127.1、KL1、L467、F1876、F2159、F1867、F852、F2266、L599、Z43961、Z43963、F854、Z43966、F915、IMS-JST002611、F240、F18、F117、CTS5820、F13、F11、F632、F110、F17、F377、F856、F1095、F1418、F1495、CTS7501、F793、Y20951、Y20932、F38、F12、F196、F930、F2685、F1365、F3232、Y15976、Y26383、FGC54486、CTS12877、F2527、F723、F1422、CTS2154、SK1691、PH203、F449、F238、F134、F1273、F724、F1266、CTS498、FGC3750、P201、M188、F2588、CTS445、CTS201、M159、M7、F2680、F1276、
CTS6489、F1275、M113、N5、Z25400、F1863、F1134、F1262、Y26403、F1837、F862、F879、F1226、F2859、P164、M134、F450、M117、M133、F42、F438、Y17728、F155、F813、Y20928、F1754、F2137、F1442、F1123、F1369、F310、F402、F1531、Z25853、CTS5492、CTS6987、Z42620、CTS4658、Z25928、SK1730、Z26010、A9462、CTS7634、F317、F3039、Y29861、CTS5488、CTS10738、
CTS9678、Z39663、A9457、YP4864、Z44068、F5524、F5525、Z44071、Z44091、CTS4960、F122、F114、F444、F79、F629、F46、FGC16847、F48、F2505、F242、CTS4266、Z26108、F2887、F3607、F3525、CTS3763、A9472、FGC16863、L1360、FGC16889、SK1768、F4249、FGC23868、CTS335、CTS53、CTS6373、A9473、F3386、Y29828、FGC34973、F1739、F743、F748、AM01822、N6、AM01856、
N7、F4110、F4068、F4124、AM01845、F717、A16433、F742、F1150、F837、F1025、F1055、F3021、P295、M45、M242、F903、M346、M3、M207、M173、M420、SRY10831.2、M17、Z645、M458、Z91、Z93、F3105、Y7、Z2124、Z2123、S4576、F1345、M343、L754、L389、P297、M269、M479、M124、FGC21706、
B254和M184；所述扩增引物的核苷酸序列为SEQ ID No .1～746所示。
2.一种包括权利要求1所述引物组合物的基于二代测序技术检测Y-SNP单倍群的试剂盒，其特征在于，还包括多重PCR反应所需的其他试剂、接头序列PCR反应所需的试剂和磁珠纯化所需要的的试剂。
3.根据权利要求2所述的试剂盒，其特征在于，检测的样本来源于血液、精液、唾液、骨骼、毛发、人体组织，然后采用血液、精液、唾液、骨骼、毛发或人体组织基因组DNA进行多重PCR。
4.一种采用如权利要求2或3所述试剂盒的基于二代测序技术检测Y-SNP单倍群的方法，其特征在于，包括如下步骤：
步骤一，多重PCR反应：以稀释好的gDNA为模板，并配制扩增反应体系进行多重片段扩增；
步骤二，对PCR扩增产物进行磁珠纯化，然后进行接头序列PCR反应；
步骤三，对步骤二的扩增产物进行磁珠纯化构建成文库；然后对文库进行浓度、片段长度和纯度进行测量；
步骤四，将文库混合、变性后加入Illumina MiSeq测序平台进行测序；将测序结果与参考基因组序列比对，并寻找SNP和Indel，然后运用ANNOVAR软件对SNP、Indel位点进行注释、确定突变位点对应的基因信息。
5.根据权利要求4所述的方法，其特征在于，步骤一的多重PCR反应体系为25μL，包括10μL ddH2O、4μL Enhancer buffer JP、4μL引物混合物、1μL gDNA以及6μL IGT-EM101polymerase mixture。
6.根据权利要求4所述的方法，其特征在于，步骤一的多重PCR反应条件为：预热105℃；95℃ 3分钟30秒；14个循环：98℃ 20秒，60℃ 3分30秒；72℃ 5分钟。
7.根据权利要求4所述的方法，其特征在于，所述接头序列PCR反应体系为25μL，包括17μL PCR磁珠纯化的产物，1μL IGT-I5 Ind ex、1μL IGT-I7 Ind ex和6μL IGT-EM101polymerase mixture。
8.根据权利要求4所述的方法，其特征在于，所述接头序列PCR反应条件为：预热105℃；95℃ 3分钟30秒；9个循环：98℃ 20秒，58℃ 1分钟，72℃ 30秒；72℃ 5分钟。
9.如权利要求1所述的引物组合物或者如权利要求2或3所述的试剂盒在家系排查和对未知样本地理来源的父系族源推断中的应用。
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