本發(fā)明屬于生物
技術領域：
，尤其涉及一種基于crispr/cas9的捕獲基因組目標序列的方法及其在高通量測序中的應用。
背景技術：
：crlspr/cas(clusteredregularlyinterspaccdshortpalindromicrepeats/crlspr-associatednuclease)技術是2013年開發(fā)的一種新的dna靶向編輯工具，被美國科學雜志評為2013年十大科學突破技術之一。cas9核酸酶，s.pyogenes，是rna介導的核酸酶，可以催化雙鏈dna特異位點切割。切割位點位于nggpam(protospaceradjacentmotif)上游序列的3個堿基處。pam序列的ngg必須連在靶點之后，位于與grna互補的dna的鏈上。其dna特異識別是靠一小段發(fā)夾rna結(jié)構(gòu)實現(xiàn)的，而dna剪切則是憑借結(jié)合于rna上的cas核酸酶執(zhí)行。crlspr序列是存在于微生物基因組中，由長度25-50bp的重復序列(repeat，r)和間區(qū)序列(spacer，s)間隔成簇排列(r-s結(jié)構(gòu))而成的dna序列，而cas則是存在于crispr簇側(cè)翼序列附近的多態(tài)性家族基因，編碼可與crispr區(qū)域發(fā)生作用的一組功能域蛋白(具有核酸酶、解旋酶、整合酶和聚合酶等的活性)。在指導rna(guiderna)的引導下對靶位點進行切割。此外，在crispr位點的第一個重復序列上游有crispr前導序列l(wèi)(leader)。該前導序列可以作為啟動子，啟動crispr序列的轉(zhuǎn)錄。前導序列、r-s結(jié)構(gòu)以及一系列的cas組成了完整的crispr/cas系統(tǒng)。人們發(fā)現(xiàn)這些crispr序列能與病毒或質(zhì)粒dna序列相匹配，表明crispr/cas系統(tǒng)能夠編碼“適應性”免疫系統(tǒng)，在細菌的獲得性免疫中發(fā)揮作用。自人類基因組圖譜繪制完成后，各國政府都加大了對人類基因組研究的扶持力度，在這樣的大背景下，基因測序技術得到了迅猛發(fā)展，基因測序市場也快速擴大，并已經(jīng)從實驗室走入臨床?；谝合嗖东@高通量測序技術，市場主要有安捷倫公司的全外顯子捕獲試劑盒，羅氏公司的固相雜交法和液相雜交法兩種技術，這些產(chǎn)品捕獲的dna序列雖然只是全基因組的部分區(qū)域，相對于全基因組測序來說成本和時間花費上具有很大的優(yōu)勢，但每個樣本的價格仍在數(shù)千元以上，超出了普通民眾的承受范圍，不適合大范圍內(nèi)篩查單基因遺傳疾病的檢測。而且這些技術的缺點是需要合成大量長片段(150－200bp)的dna，經(jīng)過pcr擴增，轉(zhuǎn)錄成加生物素修飾的rna。技術實現(xiàn)要素：本發(fā)明的一個目的是提供一種基于crispr/cas9系統(tǒng)從基因組dna中捕獲目標基因序列的方法。本發(fā)明提供的方法，包括如下步驟：1)、根據(jù)目標基因序列設計合成多個grna；每個所述grna能結(jié)合且識別所述目標基因序列不同靶區(qū)域；所述目標基因序列能通過所述grna引導cas9酶將其分隔為多個100-300bp的片段化產(chǎn)物；2)將用所述多個grna和所述cas9酶對所述基因組dna進行切割反應，得到含有多個100-300bp的片段化產(chǎn)物的切割產(chǎn)物；3)捕獲所述切割產(chǎn)物中的目標基因序列的多個100-300bp的片段化產(chǎn)物。上述切割反應僅切割不修復。上述目標基因可以是一個目標基因或多個目標基因。上述方法中，步驟1)中，所述設計合成多個grna的方法為如下a或b：a所示的方法包括如下步驟：(1)根據(jù)所述目標基因序列設計多個所述grna，再根據(jù)所述grna設計合成用于制備其對應轉(zhuǎn)錄模板的引物對，得到多個所述grna轉(zhuǎn)錄模板對應的引物對；(2)將多個所述grna轉(zhuǎn)錄模板對應的引物對分別進行聚合酶鏈式反應，得到對應的多個所述grna轉(zhuǎn)錄模板，(3)轉(zhuǎn)錄所述對應的多個grna轉(zhuǎn)錄模板，得到多個grna；b所示的方法包括如下步驟：(1)根據(jù)所述目標基因序列設計多個所述grna，再根據(jù)所述grna設計合成對應的轉(zhuǎn)錄模板，得到多個grna轉(zhuǎn)錄模板；(2)轉(zhuǎn)錄所述多個grna轉(zhuǎn)錄模板，得到多個grna。上述方法中，所述轉(zhuǎn)錄多個grna轉(zhuǎn)錄模板為分別轉(zhuǎn)錄或者混合多個grna轉(zhuǎn)錄模板再共同轉(zhuǎn)錄；或所述捕獲的方法為磁珠法。上述方法中，所述基因組dna為人或動物或其他物種的細胞的基因組dna；所述目標基因序列為brca1基因的外顯子和brca2基因的外顯子；所述brca1基因的外顯子對應的多個grna的核苷酸序列分別為序列158-220；所述brca2基因的外顯子對應的多個grna的核苷酸序列分別為序列221-316；或所述brca1基因的外顯子對應的多個grna對應的引物對中的上游引物的核苷酸序列分別為序列1-序列63，下游引物的核苷酸序列為序列157；或所述brca2基因的外顯子對應的多個grna對應的引物對中的上游引物的核苷酸序列分別為序列64-序列156，下游引物的核苷酸序列為序列157。上述的方法在目標基因序列的高通量測序文庫制備中的應用也是本發(fā)明保護的范圍。上述的方法在目標基因序列的高通量測序中的應用也是本發(fā)明保護的范圍。本發(fā)明另一個目的是提供一種目標基因序列的高通量測序方法。本發(fā)明提供的方法，包括如下步驟：1)上述的方法捕獲所述目標基因序列的多個100-300bp的片段化產(chǎn)物；2)用所述多個100-300bp的片段化產(chǎn)物制備高通量測序文庫；3)高通量測序所述高通量測序文庫，實現(xiàn)目標基因序列的高通量測序。crispr/cas9系統(tǒng)在基因組dna中捕獲或分離或聚集目標基因序列中的應用也是本發(fā)明保護的范圍；所述crispr/cas9系統(tǒng)包括多個grna和cas9酶；所述多個grna引導所述cas9酶將所述目標基因序列分割為多個大小為100-300bp片段化產(chǎn)物。本發(fā)明第3個目的是提供一種用于基因組dna中捕獲目標基因序列的試劑盒。本發(fā)明提供的試劑盒，包括上述應用中的所述crispr/cas9系統(tǒng)。本發(fā)明第4個目的是提供一種高通量測序目標基因序列的試劑盒。本發(fā)明提供的試劑盒，包括上述應用中的所述crispr/cas9系統(tǒng)和測序文庫制備所需試劑。本發(fā)明通過基于crispr基因編輯捕獲靶序列，可用于高通量測序，適用于有家族遺傳史的特定人群的篩查和大眾體檢市場；更有意義的是，本項技術還可用于開發(fā)其他單基因、多基因遺傳疾病的檢測試劑盒及外顯子測序捕獲試劑,以供臨床應用。附圖說明圖1為brca1grna和brca2grna質(zhì)檢結(jié)果。圖2為文庫擴增產(chǎn)物2％瓊脂糖凝膠電泳圖。圖3為文庫膠回收產(chǎn)物2％瓊脂糖凝膠電泳圖。圖4為文庫檢測pcr擴增產(chǎn)物2％瓊脂糖凝膠電泳圖。具體實施方式下述實施例中所使用的實驗方法如無特殊說明，均為常規(guī)方法。下述實施例中所用的材料、試劑等，如無特殊說明，均可從商業(yè)途徑得到。實施例1、基于crispr/cas9的捕獲基因組中目標基因序列的方法下面的實施例以人293t細胞基因組dna中的brca1和brca2基因為例，根據(jù)brca1基因(gi:262359905)和brca2基因(gi:568815585)的外顯子為目標基因序列。一、轉(zhuǎn)錄rna模板制備1、轉(zhuǎn)錄rna模板的引物設計合成根據(jù)目標基因組中brca1基因(gi:262359905)及brca2基因(gi:568815585)的外顯子，分別設計63個brca1grna(序列158-220)和96個brca2grna(序列221-313)，這些grna可以引導cas9酶將brca1的外顯子和brca2的外顯子全部分割為多個大小為100-300bp片段化產(chǎn)物。根據(jù)63個brca1grna和96個brca2grna分別設計63個brca1的外顯子轉(zhuǎn)錄rna模板的擴增引物和93個brca2的外顯子轉(zhuǎn)錄rna模板的擴增引物如下：用于合成63個brca1的外顯子轉(zhuǎn)錄rna模板的擴增引物：上游引物分別為brca1grna1_f-brca1grna63_f(序列1-序列63)，下游引物為grna_r(序列157)；用于合成63個brca1的外顯子轉(zhuǎn)錄rna模板的擴增引物：上游引物分別為brca2grna1_f-brca2grna93_f(序列64-序列156)，下游引物為grna_r(序列157)。2、轉(zhuǎn)錄rna模板的制備將上述brca1grna1_f-brca1grna63_f分別與下游引物grna_r在如下表1的體系中退火聚合酶鏈式反應，得到63個brca1grna1-brca1grna63的外顯子轉(zhuǎn)錄rna的模板；將上述brca2grna1_f-brca2grna93_f分別與下游引物grna_r在如下表1的體系聚合酶鏈式反應，得到93個brca2grna1-brca2grna93的外顯子轉(zhuǎn)錄rna的模板。表1為退火pcr擴增反應體系2×pfumastermixwithdye，蘇州吉瑪基因股份有限公司,目錄號：j09002。將表1的體系進行聚合酶鏈式反應，得到用以轉(zhuǎn)錄rna模板的pcr擴增產(chǎn)物。上述反應程序為：94℃3min；94℃30s，60℃30s，72℃30s，15個循環(huán)；72℃10min；4℃保存。聚合酶鏈式反應后取5μlpcr產(chǎn)物用2％瓊脂糖凝膠電泳，檢測，無非特異性條帶后，用pcr產(chǎn)物純化試劑盒(離心柱)進行純化，得到63個brca1grna-brca1grna63轉(zhuǎn)錄rna的模板和93個brca2grnabrca2grna93轉(zhuǎn)錄rna的模板。二、grna的轉(zhuǎn)錄將上述63個brca1grna1-brca1grna63的轉(zhuǎn)錄rna模板和93個brca2grna1–brca2grna93的轉(zhuǎn)錄rna模板按照等質(zhì)量比混合，按照transcriptaidt7highyieldtranscriptionkit(賽默飛世爾科技(中國)有限公司；目錄號：k0441)說明書共同轉(zhuǎn)錄，得到63個brca1grna1-brca1grna63和93個brca2grna1–brca2grna93的混合物。上述轉(zhuǎn)錄體系如表2所示，表2為轉(zhuǎn)錄反應體系組份不含核酸酶的水to20μl5×transcriptaidreactionbuffer4μlatp/ctp/gtp/utpmix*8μl轉(zhuǎn)錄rna模板2μg**transcriptaidenzymemix2μl總體積20μl37℃轉(zhuǎn)錄4小時，得到轉(zhuǎn)錄產(chǎn)物。將轉(zhuǎn)錄產(chǎn)物按照如下方法純化，得到63個brca1grna-brca1grna63和93個brca2grna–brca2grna93的混合物：(1)轉(zhuǎn)錄產(chǎn)物加入2μldnasei混合，37℃反應35分鐘；(2)20μl反應產(chǎn)物中加入115μl無核酸酶的水，15μl3m醋酸鈉；(3)加入等體積的水飽和酚，氯仿各75μl，混合，12000轉(zhuǎn)/分離心5分鐘；(4)重復(3)一次；(5)加入2倍體積-20℃預冷的無水乙醇沉淀rna,-20℃放置過夜；(6)4℃12000轉(zhuǎn)/分離心30分鐘，去上清；(7)加入1ml70％乙醇，12000轉(zhuǎn)/分離心5分鐘，去上清；(8)室溫干燥，加入20μldepc水處溶解rna。將純化后63個brca1grna1-brca1grna63和93個brca2grna1–brca2grna93進行質(zhì)檢，結(jié)果如圖1所示，泳道m(xù)為10bpdnaladder泳道1和2均為200ng轉(zhuǎn)錄的grna，可以看出，已得到轉(zhuǎn)錄的grna。上述質(zhì)檢的方法如下：(1)配制15％變性膠；(2)200v預電泳30分鐘；(3)取200ng的rna加入2×rna上樣緩沖液；(4)于65℃變性5分鐘，立即冰??；(5)200v電泳60分鐘；(6)于染膠盒中染色10分鐘；(7)拍照。三、目標基因組dna體外切割1)將上述純化后63個brca1grna1-brca1grna63和93個brca2grna1–brca2grna93混合物取10μl稀釋至15μm，再90℃加熱5分鐘，緩慢冷卻至室溫(形成二級結(jié)構(gòu))，得到處理后63個brca1grna1-brca1grna63和處理后93個brca2grna1–brca2grna93。2)將處理后63個brca1grna1-brca1grna63和處理后93個brca2grna1–brca2grna93混合物加入表3所示的切割反應體系中，室溫孵育15分鐘，然后加入5.5μl100ng/μl的人293t細胞基因組dna。表3基因組dna切割反應體系volume(μl)不含核酸酶的水27.55×cleavagebuffer11.0cas9核酸酶：15μm5.5反應產(chǎn)物：15μm5.5cas9核酸酶，s.pyogenes,neb(北京)有限公司,neb(北京)有限公司,m0386m。在37℃孵育30分鐘，得到反應產(chǎn)物。(3)將2)得到的反應產(chǎn)物加入5μl500mmedta，加入22μl蛋白酶k消化cas9，室溫孵育20分鐘，得到切割產(chǎn)物。四、從切割產(chǎn)物中磁珠法序列捕獲片段化產(chǎn)物1.振蕩ampurexp磁珠(貝克曼庫爾特有限公司生產(chǎn)的agencourtampurexpkit:產(chǎn)品目錄號為a63880)，充分混勻；2.向上述三得到的切割產(chǎn)物中加入0.7×的磁珠，用移液器混勻10次，室溫放置1分鐘；3.置于磁力架上吸附5分鐘，轉(zhuǎn)移上清至新的離心管中；4.加入1.1倍體積的磁珠，用移液器混勻10次，室溫放置1分鐘；5.向磁珠中加入200μl新鮮配制的80％的乙醇，室溫放置30s，棄上清；6.重復上述步驟1次；7.開蓋室溫放置10分鐘；8.加入50μl不含核酸酶的水，用移液器混勻，室溫放置1分鐘；9.置于磁力架上5分鐘，轉(zhuǎn)移上清至新的離心管中，得到含有brca1的外顯子大小為100-300bp片段化產(chǎn)物和brca2的外顯子大小為100-300bp片段化產(chǎn)物的混合物。實施例2、高通量測序目標靶序列一、高通量測序文庫構(gòu)建1、末端修復、磷酸化并加da尾(1)在滅菌ep管中加入下列試劑表4所示：表4為末端修復、磷酸化并加da尾體系(2)使用移液器輕輕吹打混勻，短暫離心將反應液收集至管底。(3)將反應管置于pcr儀，運行表5程序，得到反應產(chǎn)物：表52、接頭連接(1)向步驟1得到的反應產(chǎn)物中加入下列組分：表6t4dna連接酶,neb(北京)有限公司,neb(北京)有限公司,m0202l。y型接頭(uaf/ai1)uafaatgatacggcgaccaccgagatctacactctttccctacacgacgctcttccgatc*tai1gatcggaagagcacacgtctgaactccagtcacatcacgatctcgtatgccgtcttctgcttg*硫代修飾，由上海捷瑞生物工程有限公司合成。(2)使用移液器輕輕吹打混勻，短暫離心將反應液收集至管底。(3)將反應管置于pcr儀，運行下列程序，得到接頭連接產(chǎn)物：表73、接頭連接產(chǎn)物純化和大小分選(1)渦旋振蕩混勻ampurexpbeads(可為貝克曼庫爾特有限公司生產(chǎn)的agencourtampurexpkit:產(chǎn)品目錄號為a63880)；(2)向50ul上述2得到的接頭連接產(chǎn)物入50ulampurexpbeads；(3)用移液器混勻10次，室溫放置5min；(4)將磁珠置于磁力架吸附2min，棄掉上清；(5)向磁珠加入200ul新鮮80％乙醇漂洗磁珠，室溫放置30sec,磁鐵吸附去上清；(6)重復上步驟1次；(7)開蓋放置10min；(8)從磁鐵上取下磁珠加入50uldepc水，用移液器混勻10次；(9)將磁珠置于磁鐵～5min；(10)取50ul上清至一個新的ep管中；(11)渦旋振蕩混勻ampurexpbeads；(12)向上步50ul洗脫產(chǎn)物入50ulampurexpbeads；(13)用移液器混勻10次，室溫放置5min；(14)將磁珠置于磁力架吸附2min，棄上清；(15)向磁珠加入200ul新鮮80％乙醇漂洗磁珠，室溫放置30sec,磁鐵吸附去上清；(16)重復上步驟1次；(17)開蓋放置10min；(18)從磁鐵上取下磁珠加入40uldepc水，用移液器混勻10次；(19)將磁珠置于磁鐵～5min；(20)吸取約40ul上清至一個新的ep管中，得到純化后接頭連接產(chǎn)物。4、接頭連接產(chǎn)物pcr擴增(1)配制下列反應：表8引物mpfaatgatacggcgaccaccgagatctacactctttccctacacgacgctcttccgatct引物mrpi1caagcagaagacggcatacgagatcgtgatgtgactggagttcagacgtgtgctcttccgatc*t由上海捷瑞生物工程有限公司合成。kapahifidna聚合酶：kapabiosystems,目錄號：kk2102(2)使用移液器輕輕吹打混勻，短暫離心將反應液收集至管底。將反應管置于pcr儀，運行：反應程序為：94℃3min；94℃30s，60℃30s，72℃30s，15個循環(huán)；72℃10min；4℃保存。把擴增產(chǎn)物進行2％瓊脂糖凝膠電泳，結(jié)果如圖2所示，m:20bpdnaladder；1：文庫擴增產(chǎn)物，可以看出，得到文庫擴增產(chǎn)物。5、pcr擴增產(chǎn)物純化(1)切取上述4電泳中所需條帶，用上海捷瑞生物工程有限公司的瓊脂糖凝膠dna回收試劑盒(離心柱型)：gk2042-50進行凝膠收。(2)向凝膠中加入400μlbindingsolution，置于50℃水浴鍋中，至膠塊溶解。(3)期間每隔2分鐘搖動一次。(4)將溶好的膠塊轉(zhuǎn)移至硅膠柱中，室溫放置2分鐘，6000轉(zhuǎn)/分離心1分鐘，棄廢液。(5)向硅膠柱中加入500μlwashingsolution，室溫放置3分鐘。(6)12000轉(zhuǎn)/分離心1分鐘，棄廢液。(7)重復(6)一次。(8)12000轉(zhuǎn)/分離心1分鐘，轉(zhuǎn)移硅膠柱至新的1.5ml離心管中(9)向硅膠柱中加入30μl不含核酸酶的水，室溫放置2分鐘。(10)12000轉(zhuǎn)/分離心1分鐘，收集上清，得到純化后pcr擴增產(chǎn)物。將純化后pcr擴增產(chǎn)物進行電泳檢測，結(jié)果如圖3所示，m:20bpdnaladder；1：文庫膠回收產(chǎn)物，可以看出，得到高通量測序文庫。6、捕獲文庫檢測(1)pcr反應體系配制表9文庫質(zhì)檢反應體系組份體積(μl)不含核酸酶的水16.210×pcrbuffer2.5mgcl2:25mm2.510mmdntpmix0.5taqdna聚合酶0.35得到的純化后pcr擴增產(chǎn)物1總體積23μltaqdnapolymerase為promega公司的產(chǎn)品，產(chǎn)品目錄號為m1665s；根據(jù)樣品數(shù)量配制反應體系以23μl/管，分裝至0.2ml的pcr管中，分別標記為1-10,分別再向pcr管中加入不同的上下游引物(該上下游引物用于擴增大小為100-300bp片段化產(chǎn)物，每對引物對應一個100-300bp片段，隨機選取其中10對引物)，如下：表10(2)反應程序：反應程序為：94℃3min；94℃30s，60℃30s，72℃30s，22個循環(huán)；72℃10min；4℃保存(3)擴增產(chǎn)物進行2％瓊脂糖凝膠電泳，結(jié)果如圖4所示，20bpdnaladder；1：rs55906931；2：rs4986850；2：rs799917；4：rs80357280；5：rs56012641；6：rs1800704；7：rs80357420；8：rs1799950；9：rs4986852；10：rs16941；可以看出，均擴增出目的條帶，表明捕獲序列中有brca1的外顯子大小為100-300bp片段化產(chǎn)物和brca2的外顯子大小為100-300bp片段化產(chǎn)物。7、高通量測序及數(shù)據(jù)分析取步驟4構(gòu)建好的文庫用hiseq3000平臺進行測序，對測序數(shù)據(jù)用fastx-toolkit0.0.13軟件(網(wǎng)址為：http://hannonlab.cshl.edu/fastx_toolkit/)中的fastq_quality_trimmer模塊去除低質(zhì)量reads，用fastx_clipper模塊去除接頭序列，分隔含n的讀段數(shù)，提取分隔后的配對讀段；測序數(shù)據(jù)預處理后的結(jié)果見表11。表11測序數(shù)據(jù)質(zhì)量預處理數(shù)據(jù)統(tǒng)計結(jié)果使用bwa-0.7.15進行有基因參照的參考基因組mapping。從結(jié)果上看，捕獲文庫比對到參考基因組上的讀段數(shù)比例x在84％以上，其中成對兒reads比對的比例約83％，結(jié)果正常。與參考基因組比對后的統(tǒng)計結(jié)果見表12。表12參考基因組比對整體情況統(tǒng)計用bwa-0.7.15軟件(網(wǎng)址為：https://sourceforge.net/projects/bio-bwa/files/)對有效數(shù)據(jù)與基因組dna進行比對，用samtools-0.1.9軟件(網(wǎng)址為：https://sourceforge.net/projects/samtools/files/samtools/0.1.19/)把比對的sam格式轉(zhuǎn)換成bam格式，用bedtools-2.17.0軟件(網(wǎng)址為：https://github.com/arq5x/bedtools/releases/tag/v2.17.0)中的coverage模塊計算靶區(qū)域的覆蓋率。靶區(qū)域的覆蓋率見表13；表13富集效率靶序列捕獲文庫覆蓋率(％)s96.58％根據(jù)公式用teqc3.16.0軟件(網(wǎng)址為：https://bioconductor.org/packages/release/bioc/html/teqc.htm)計算靶區(qū)域的富集效率，靶區(qū)域的富集效率見表14。表14靶區(qū)域的富集效率統(tǒng)計結(jié)果上述結(jié)果表明，本發(fā)明所提供的crispr探針組合可用于構(gòu)建捕獲文庫，進一步利用該文庫捕獲基因組靶序列。因此，本發(fā)明所提供的方法，可制備靶序列的捕獲文庫。序列表<110>蘇州吉賽基因測序科技有限公司<120>一種基于crispr/cas9的捕獲基因組目標序列的方法及其在高通量測序中的應用<160>333<170>patentinversion3.5<210>1<211>58<212>dna<213>人工序列<220><223><400>1taatacgactcactataggcaaacttaggtattggaaccgttttagagctagaaatag58<210>2<211>58<212>dna<213>人工序列<220><223><400>2taatacgactcactataggttcttcacagtgcagtgaatgttttagagctagaaatag58<210>3<211>58<212>dna<213>人工序列<220><223><400>3taatacgactcactataggatataccttctcagtctactgttttagagctagaaatag58<210>4<211>58<212>dna<213>人工序列<220><223><400>4taatacgactcactatagggttagatgatggtgaaataagttttagagctagaaatag58<210>5<211>58<212>dna<213>人工序列<220><223><400>5taatacgactcactatagggaatgctatgcttagattaggttttagagctagaaatag58<210>6<211>58<212>dna<213>人工序列<220><223><400>6taatacgactcactataggtctgctagaggaaaactttggttttagagctagaaatag58<210>7<211>58<212>dna<213>人工序列<220><223><400>7taatacgactcactataggactgcaggctttcctgtggtgttttagagctagaaatag58<210>8<211>58<212>dna<213>人工序列<220><223><400>8taatacgactcactataggtcgggaaacaagcatagaaagttttagagctagaaatag58<210>9<211>58<212>dna<213>人工序列<220><223><400>9taatacgactcactataggagcagtatttcattggtaccgttttagagctagaaatag58<210>10<211>58<212>dna<213>人工序列<220><223><400>10taatacgactcactatagggaaggtaaagaacctgcaacgttttagagctagaaatag58<210>11<211>58<212>dna<213>人工序列<220><223><400>11taatacgactcactataggtgaggaggaagtcttctaccgttttagagctagaaatag58<210>12<211>58<212>dna<213>人工序列<220><223><400>12taatacgactcactataggactcctgaaatgataaatcagttttagagctagaaatag58<210>13<211>58<212>dna<213>人工序列<220><223><400>13taatacgactcactataggactggccagtgatcctcatggttttagagctagaaatag58<210>14<211>58<212>dna<213>人工序列<220><223><400>14taatacgactcactatagggagatactgaagatgttcctgttttagagctagaaatag58<210>15<211>58<212>dna<213>人工序列<220><223><400>15taatacgactcactataggagacagaatgaatgtagaaagttttagagctagaaatag58<210>16<211>58<212>dna<213>人工序列<220><223><400>16taatacgactcactataggtgcttgtgaattttctgagagttttagagctagaaatag58<210>17<211>58<212>dna<213>人工序列<220><223><400>17taatacgactcactataggcttcagtccttctactgtccgttttagagctagaaatag58<210>18<211>56<212>dna<213>人工序列<220><223><400>18taatacgactcactataggctctaatcaatcgactccgttttagagctagaaatag56<210>19<211>57<212>dna<213>人工序列<220><223><400>19taatacgactcactataggcatgtacctgtgctatatggttttagagctagaaatag57<210>20<211>58<212>dna<213>人工序列<220><223><400>20taatacgactcactataggagcattcaaagtgtcaaagtgttttagagctagaaatag58<210>21<211>58<212>dna<213>人工序列<220><223><400>21taatacgactcactataggctacagtaggggcatccatagttttagagctagaaatag58<210>22<211>58<212>dna<213>人工序列<220><223><400>22taatacgactcactataggaccctacactctccggatgagttttagagctagaaatag58<210>23<211>58<212>dna<213>人工序列<220><223><400>23taatacgactcactataggatctgaggaacccccatcgtgttttagagctagaaatag58<210>24<211>58<212>dna<213>人工序列<220><223><400>24taatacgactcactataggtccaatgtccagaacactacgttttagagctagaaatag58<210>25<211>58<212>dna<213>人工序列<220><223><400>25taatacgactcactataggagtgctagatactttcacacgttttagagctagaaatag58<210>26<211>57<212>dna<213>人工序列<220><223><400>26taatacgactcactataggagcagacacgtcatatttagttttagagctagaaatag57<210>27<211>58<212>dna<213>人工序列<220><223><400>27taatacgactcactataggaggatcatcaagaattatgcgttttagagctagaaatag58<210>28<211>58<212>dna<213>人工序列<220><223><400>28taatacgactcactataggagcgattcacaaaagagcacgttttagagctagaaatag58<210>29<211>58<212>dna<213>人工序列<220><223><400>29taatacgactcactataggcacctaacgtttaacacctagttttagagctagaaatag58<210>30<211>58<212>dna<213>人工序列<220><223><400>30taatacgactcactataggttacactcccaagatcaatcgttttagagctagaaatag58<210>31<211>58<212>dna<213>人工序列<220><223><400>31taatacgactcactataggccaccgtgcctcgcctcatggttttagagctagaaatag58<210>32<211>58<212>dna<213>人工序列<220><223><400>32taatacgactcactataggtgcacgttctacacgtgtccgttttagagctagaaatag58<210>33<211>57<212>dna<213>人工序列<220><223><400>33taatacgactcactataggttgttaagtcttagtcattgttttagagctagaaatag57<210>34<211>57<212>dna<213>人工序列<220><223><400>34taatacgactcactataggccccagagtcagctcgtgtgttttagagctagaaatag57<210>35<211>58<212>dna<213>人工序列<220><223><400>35taatacgactcactataggtgggtacatgaatacagtgtgttttagagctagaaatag58<210>36<211>58<212>dna<213>人工序列<220><223><400>36taatacgactcactataggttagatgtactagtctatcagttttagagctagaaatag58<210>37<211>58<212>dna<213>人工序列<220><223><400>37taatacgactcactataggaccacctatcatctaatgatgttttagagctagaaatag58<210>38<211>57<212>dna<213>人工序列<220><223><400>38taatacgactcactataggtcaaccctgacatattggcgttttagagctagaaatag57<210>39<211>58<212>dna<213>人工序列<220><223><400>39taatacgactcactataggcatgttgtagcttatgttatgttttagagctagaaatag58<210>40<211>58<212>dna<213>人工序列<220><223><400>40taatacgactcactataggcttctggattctggcttatagttttagagctagaaatag58<210>41<211>58<212>dna<213>人工序列<220><223><400>41taatacgactcactataggagacagatgctagcaccaaagttttagagctagaaatag58<210>42<211>58<212>dna<213>人工序列<220><223><400>42taatacgactcactataggacaactaatataccagtcaggttttagagctagaaatag58<210>43<211>58<212>dna<213>人工序列<220><223><400>43taatacgactcactataggtgctttgttctggatttcgcgttttagagctagaaatag58<210>44<211>56<212>dna<213>人工序列<220><223><400>44taatacgactcactataggaattgaaatcacctagtagttttagagctagaaatag56<210>45<211>58<212>dna<213>人工序列<220><223><400>45taatacgactcactataggaggtagtatgagttccatcagttttagagctagaaatag58<210>46<211>58<212>dna<213>人工序列<220><223><400>46taatacgactcactataggtttaaggtgaagcagcatctgttttagagctagaaatag58<210>47<211>58<212>dna<213>人工序列<220><223><400>47taatacgactcactataggattactggtggacttacttcgttttagagctagaaatag58<210>48<211>58<212>dna<213>人工序列<220><223><400>48taatacgactcactataggctctgtcaaatgtcgtggtagttttagagctagaaatag58<210>49<211>58<212>dna<213>人工序列<220><223><400>49taatacgactcactataggttcttgatctcccacactatgttttagagctagaaatag58<210>50<211>57<212>dna<213>人工序列<220><223><400>50taatacgactcactataggaaatggttgctgggcacgggttttagagctagaaatag57<210>51<211>58<212>dna<213>人工序列<220><223><400>51taatacgactcactataggtataggtaaacacacgacttgttttagagctagaaatag58<210>52<211>58<212>dna<213>人工序列<220><223><400>52taatacgactcactataggttctgaagataccgttaatagttttagagctagaaatag58<210>53<211>58<212>dna<213>人工序列<220><223><400>53taatacgactcactataggttaaaggcatgggcttcgccgttttagagctagaaatag58<210>54<211>58<212>dna<213>人工序列<220><223><400>54taatacgactcactataggagtgaattttatgagccctagttttagagctagaaatag58<210>55<211>57<212>dna<213>人工序列<220><223><400>55taatacgactcactataggacgtctgtctacattgaatgttttagagctagaaatag57<210>56<211>58<212>dna<213>人工序列<220><223><400>56taatacgactcactataggcagctaacatgtatgatgccgttttagagctagaaatag58<210>57<211>57<212>dna<213>人工序列<220><223><400>57taatacgactcactataggccttagcccccttagtagcgttttagagctagaaatag57<210>58<211>56<212>dna<213>人工序列<220><223><400>58taatacgactcactataggtttctgtagcccatacttgttttagagctagaaatag56<210>59<211>58<212>dna<213>人工序列<220><223><400>59taatacgactcactataggagcatacatagggtttctctgttttagagctagaaatag58<210>60<211>58<212>dna<213>人工序列<220><223><400>60taatacgactcactataggcagcacttgagtgtcattctgttttagagctagaaatag58<210>61<211>58<212>dna<213>人工序列<220><223><400>61taatacgactcactataggcatctgtataaaccgtgtgagttttagagctagaaatag58<210>62<211>58<212>dna<213>人工序列<220><223><400>62taatacgactcactataggatttggtaatgatgctaggtgttttagagctagaaatag58<210>63<211>57<212>dna<213>人工序列<220><223><400>63taatacgactcactataggccatttaaaaagtaatggcgttttagagctagaaatag57<210>64<211>58<212>dna<213>人工序列<220><223><400>64taatacgactcactataggaagcattggaggaatatcgtgttttagagctagaaatag58<210>65<211>58<212>dna<213>人工序列<220><223><400>65taatacgactcactataggacaacataatcatcgtttgcgttttagagctagaaatag58<210>66<211>57<212>dna<213>人工序列<220><223><400>66taatacgactcactataggtcactggttaaaactaagggttttagagctagaaatag57<210>67<211>58<212>dna<213>人工序列<220><223><400>67taatacgactcactataggagtaatgcaatatggtagacgttttagagctagaaatag58<210>68<211>58<212>dna<213>人工序列<220><223><400>68taatacgactcactataggcagtgagaatgtatatactcgttttagagctagaaatag58<210>69<211>58<212>dna<213>人工序列<220><223><400>69taatacgactcactataggtaatattttggctaagagccgttttagagctagaaatag58<210>70<211>58<212>dna<213>人工序列<220><223><400>70taatacgactcactataggttctcattcatataaattgtgttttagagctagaaatag58<210>71<211>58<212>dna<213>人工序列<220><223><400>71taatacgactcactataggttttacccccagtggtatgtgttttagagctagaaatag58<210>72<211>58<212>dna<213>人工序列<220><223><400>72taatacgactcactataggtagaaatgccctgatcattagttttagagctagaaatag58<210>73<211>58<212>dna<213>人工序列<220><223><400>73taatacgactcactataggctatgagcacagtagaactagttttagagctagaaatag58<210>74<211>58<212>dna<213>人工序列<220><223><400>74taatacgactcactataggtacatttagtggtagtccaggttttagagctagaaatag58<210>75<211>58<212>dna<213>人工序列<220><223><400>75taatacgactcactataggcttaagtatagtaattagaagttttagagctagaaatag58<210>76<211>57<212>dna<213>人工序列<220><223><400>76taatacgactcactataggcttagataaattacagattgttttagagctagaaatag57<210>77<211>58<212>dna<213>人工序列<220><223><400>77taatacgactcactataggtagtagtccccccttatccagttttagagctagaaatag58<210>78<211>58<212>dna<213>人工序列<220><223><400>78taatacgactcactataggttgcggtaaaccgagatcacgttttagagctagaaatag58<210>79<211>58<212>dna<213>人工序列<220><223><400>79taatacgactcactataggtcaggctttactagaagaacgttttagagctagaaatag58<210>80<211>58<212>dna<213>人工序列<220><223><400>80taatacgactcactataggaatagctgcaaagaccacatgttttagagctagaaatag58<210>81<211>58<212>dna<213>人工序列<220><223><400>81taatacgactcactataggattcatcagcgtttgcttcagttttagagctagaaatag58<210>82<211>58<212>dna<213>人工序列<220><223><400>82taatacgactcactataggcaaggaagttgtaccgtcttgttttagagctagaaatag58<210>83<211>57<212>dna<213>人工序列<220><223><400>83taatacgactcactataggcagtaaagcaggcaatatcgttttagagctagaaatag57<210>84<211>58<212>dna<213>人工序列<220><223><400>84taatacgactcactataggtctgtagctttgaagaatgcgttttagagctagaaatag58<210>85<211>58<212>dna<213>人工序列<220><223><400>85taatacgactcactataggacatttgcaaatgctgattcgttttagagctagaaatag58<210>86<211>58<212>dna<213>人工序列<220><223><400>86taatacgactcactataggagtgacctgattctaaacacgttttagagctagaaatag58<210>87<211>58<212>dna<213>人工序列<220><223><400>87taatacgactcactataggccaggagttcgagactagccgttttagagctagaaatag58<210>88<211>58<212>dna<213>人工序列<220><223><400>88taatacgactcactataggtgtcccaaaagagctagttagttttagagctagaaatag58<210>89<211>58<212>dna<213>人工序列<220><223><400>89taatacgactcactataggtccacttttgaatgttgtacgttttagagctagaaatag58<210>90<211>58<212>dna<213>人工序列<220><223><400>90taatacgactcactataggaaaatgtcagacaagctcaagttttagagctagaaatag58<210>91<211>58<212>dna<213>人工序列<220><223><400>91taatacgactcactataggtcttccaagtagctaatgaagttttagagctagaaatag58<210>92<211>57<212>dna<213>人工序列<220><223><400>92taatacgactcactataggattacatgaacaaatgggcgttttagagctagaaatag57<210>93<211>57<212>dna<213>人工序列<220><223><400>93taatacgactcactataggatacagtattaattgactggttttagagctagaaatag57<210>94<211>58<212>dna<213>人工序列<220><223><400>94taatacgactcactataggtggtctttaagatagtcatcgttttagagctagaaatag58<210>95<211>58<212>dna<213>人工序列<220><223><400>95taatacgactcactataggaggggcttttattctgctcagttttagagctagaaatag58<210>96<211>58<212>dna<213>人工序列<220><223><400>96taatacgactcactataggaataatattgaaatgactacgttttagagctagaaatag58<210>97<211>58<212>dna<213>人工序列<220><223><400>97taatacgactcactataggattatctggccagtttatgagttttagagctagaaatag58<210>98<211>57<212>dna<213>人工序列<220><223><400>98taatacgactcactataggaaaagttatgcaattcttcgttttagagctagaaatag57<210>99<211>58<212>dna<213>人工序列<220><223><400>99taatacgactcactataggtgggttttcatacagctagcgttttagagctagaaatag58<210>100<211>58<212>dna<213>人工序列<220><223><400>100taatacgactcactataggttatcacttaagagcttagggttttagagctagaaatag58<210>101<211>58<212>dna<213>人工序列<220><223><400>101taatacgactcactataggacaacttgtgtaaaaagctagttttagagctagaaatag58<210>102<211>58<212>dna<213>人工序列<220><223><400>102taatacgactcactataggatatacctcatcagaatggtgttttagagctagaaatag58<210>103<211>58<212>dna<213>人工序列<220><223><400>103taatacgactcactataggcctgcatttaggatagccaggttttagagctagaaatag58<210>104<211>58<212>dna<213>人工序列<220><223><400>104taatacgactcactataggatgtagcacgcattcacatagttttagagctagaaatag58<210>105<211>56<212>dna<213>人工序列<220><223><400>105taatacgactcactatagggatttttagcacagcaaggttttagagctagaaatag56<210>106<211>57<212>dna<213>人工序列<220><223><400>106taatacgactcactataggtaatgaagcatctgataccgttttagagctagaaatag57<210>107<211>58<212>dna<213>人工序列<220><223><400>107taatacgactcactataggcaaagttaagggagtgttaggttttagagctagaaatag58<210>108<211>57<212>dna<213>人工序列<220><223><400>108taatacgactcactataggacaaacaacagttggtattgttttagagctagaaatag57<210>109<211>58<212>dna<213>人工序列<220><223><400>109taatacgactcactataggagagaatgtgtggcatgactgttttagagctagaaatag58<210>110<211>58<212>dna<213>人工序列<220><223><400>110taatacgactcactataggcgtatacagatttgatatctgttttagagctagaaatag58<210>111<211>58<212>dna<213>人工序列<220><223><400>111taatacgactcactataggactttactctttcaaacattgttttagagctagaaatag58<210>112<211>58<212>dna<213>人工序列<220><223><400>112taatacgactcactataggctgttctccctctataggtagttttagagctagaaatag58<210>113<211>58<212>dna<213>人工序列<220><223><400>113taatacgactcactataggtgcttgtactgtgagttattgttttagagctagaaatag58<210>114<211>58<212>dna<213>人工序列<220><223><400>114taatacgactcactataggtttaaacatgtcttaccgaagttttagagctagaaatag58<210>115<211>57<212>dna<213>人工序列<220><223><400>115taatacgactcactataggatgaagtactaaggttgaggttttagagctagaaatag57<210>116<211>57<212>dna<213>人工序列<220><223><400>116taatacgactcactataggcagaccctcatttgctacagttttagagctagaaatag57<210>117<211>58<212>dna<213>人工序列<220><223><400>117taatacgactcactataggatgagacacttgattactacgttttagagctagaaatag58<210>118<211>58<212>dna<213>人工序列<220><223><400>118taatacgactcactataggagttacagctactgcttgatgttttagagctagaaatag58<210>119<211>58<212>dna<213>人工序列<220><223><400>119taatacgactcactataggcaatctaggactgctgttacgttttagagctagaaatag58<210>120<211>58<212>dna<213>人工序列<220><223><400>120taatacgactcactataggtgaggcgggcagatcatctggttttagagctagaaatag58<210>121<211>58<212>dna<213>人工序列<220><223><400>121taatacgactcactataggctctgcgtgttctcataaacgttttagagctagaaatag58<210>122<211>58<212>dna<213>人工序列<220><223><400>122taatacgactcactataggccccttgctaggcctgcctcgttttagagctagaaatag58<210>123<211>58<212>dna<213>人工序列<220><223><400>123taatacgactcactataggtagcaggaggcgtataaacggttttagagctagaaatag58<210>124<211>58<212>dna<213>人工序列<220><223><400>124taatacgactcactataggtttgtgtagctgtatacgtagttttagagctagaaatag58<210>125<211>58<212>dna<213>人工序列<220><223><400>125taatacgactcactataggtgtctctcgaactaaaaagtgttttagagctagaaatag58<210>126<211>57<212>dna<213>人工序列<220><223><400>126taatacgactcactataggaattcagtatcatcctatggttttagagctagaaatag57<210>127<211>58<212>dna<213>人工序列<220><223><400>127taatacgactcactataggcgtaatcatatacggcagtagttttagagctagaaatag58<210>128<211>58<212>dna<213>人工序列<220><223><400>128taatacgactcactataggtattagcaatccccaaataggttttagagctagaaatag58<210>129<211>58<212>dna<213>人工序列<220><223><400>129taatacgactcactataggatcggctataaaaaagataagttttagagctagaaatag58<210>130<211>58<212>dna<213>人工序列<220><223><400>130taatacgactcactataggtggggcttcaagaggtgtacgttttagagctagaaatag58<210>131<211>58<212>dna<213>人工序列<220><223><400>131taatacgactcactataggttaagctcaagaaagatctcgttttagagctagaaatag58<210>132<211>57<212>dna<213>人工序列<220><223><400>132taatacgactcactataggcttcctcaaaaaatctacagttttagagctagaaatag57<210>133<211>58<212>dna<213>人工序列<220><223><400>133taatacgactcactataggagtttggtataccagcgagcgttttagagctagaaatag58<210>134<211>57<212>dna<213>人工序列<220><223><400>134taatacgactcactataggagaatcacttgaacccggggttttagagctagaaatag57<210>135<211>58<212>dna<213>人工序列<220><223><400>135taatacgactcactataggcaagttaattgtatcaggccgttttagagctagaaatag58<210>136<211>58<212>dna<213>人工序列<220><223><400>136taatacgactcactataggagccttattcactaaaattcgttttagagctagaaatag58<210>137<211>57<212>dna<213>人工序列<220><223><400>137taatacgactcactataggtatctagggtattctttttgttttagagctagaaatag57<210>138<211>58<212>dna<213>人工序列<220><223><400>138taatacgactcactataggtctcactctgtcacccaatcgttttagagctagaaatag58<210>139<211>58<212>dna<213>人工序列<220><223><400>139taatacgactcactataggtgtcttgttagtgcacgtgagttttagagctagaaatag58<210>140<211>58<212>dna<213>人工序列<220><223><400>140taatacgactcactataggttcaactatataccgagtaggttttagagctagaaatag58<210>141<211>58<212>dna<213>人工序列<220><223><400>141taatacgactcactataggtagagctcatcttaagggtggttttagagctagaaatag58<210>142<211>58<212>dna<213>人工序列<220><223><400>142taatacgactcactataggcaagggatgtcacaaccgtggttttagagctagaaatag58<210>143<211>58<212>dna<213>人工序列<220><223><400>143taatacgactcactataggaaacagttatactgagtattgttttagagctagaaatag58<210>144<211>58<212>dna<213>人工序列<220><223><400>144taatacgactcactataggagtgaaggggctcccgtggcgttttagagctagaaatag58<210>145<211>57<212>dna<213>人工序列<220><223><400>145taatacgactcactataggattagttggagctaccagtgttttagagctagaaatag57<210>146<211>57<212>dna<213>人工序列<220><223><400>146taatacgactcactataggctgaatttggcgaaagctcgttttagagctagaaatag57<210>147<211>58<212>dna<213>人工序列<220><223><400>147taatacgactcactataggatttggattctggtcgccacgttttagagctagaaatag58<210>148<211>58<212>dna<213>人工序列<220><223><400>148taatacgactcactataggccccatctcctgaggttcatgttttagagctagaaatag58<210>149<211>57<212>dna<213>人工序列<220><223><400>149taatacgactcactataggcatcggcatgtttgacaatgttttagagctagaaatag57<210>150<211>58<212>dna<213>人工序列<220><223><400>150taatacgactcactataggttagttggggtggaccacttgttttagagctagaaatag58<210>151<211>57<212>dna<213>人工序列<220><223><400>151taatacgactcactataggctcaaatcattcctggtacgttttagagctagaaatag57<210>152<211>58<212>dna<213>人工序列<220><223><400>152taatacgactcactataggtgaagcaagatatgaaactcgttttagagctagaaatag58<210>153<211>58<212>dna<213>人工序列<220><223><400>153taatacgactcactataggcctaacctattaggagttaggttttagagctagaaatag58<210>154<211>58<212>dna<213>人工序列<220><223><400>154taatacgactcactataggcagtctactcaagaaatccagttttagagctagaaatag58<210>155<211>57<212>dna<213>人工序列<220><223><400>155taatacgactcactataggctcttttgtctggttcaacgttttagagctagaaatag57<210>156<211>57<212>dna<213>人工序列<220><223><400>156taatacgactcactataggcaacataagtactaatgtggttttagagctagaaatag57<210>157<211>78<212>dna<213>人工序列<220><223><400>157aagcaccgactcggtgccactttttcaagttgataacggactagccttattttaacttgc60tatttctagctctaaaac78<210>158<211>41<212>dna<213>人工序列<220><223><400>158ggcaaacuuagguauuggaaccguuuuagagcuagaaauag41<210>159<211>41<212>dna<213>人工序列<220><223><400>159gguucuucacagugcagugaauguuuuagagcuagaaauag41<210>160<211>41<212>dna<213>人工序列<220><223><400>160ggauauaccuucucagucuacuguuuuagagcuagaaauag41<210>161<211>41<212>dna<213>人工序列<220><223><400>161ggguuagaugauggugaaauaaguuuuagagcuagaaauag41<210>162<211>41<212>dna<213>人工序列<220><223><400>162gggaaugcuaugcuuagauuagguuuuagagcuagaaauag41<210>163<211>41<212>dna<213>人工序列<220><223><400>163ggucugcuagaggaaaacuuugguuuuagagcuagaaauag41<210>164<211>41<212>dna<213>人工序列<220><223><400>164ggacugcaggcuuuccugugguguuuuagagcuagaaauag41<210>165<211>41<212>dna<213>人工序列<220><223><400>165ggucgggaaacaagcauagaaaguuuuagagcuagaaauag41<210>166<211>41<212>dna<213>人工序列<220><223><400>166ggagcaguauuucauugguaccguuuuagagcuagaaauag41<210>167<211>41<212>dna<213>人工序列<220><223><400>167gggaagguaaagaaccugcaacguuuuagagcuagaaauag41<210>168<211>41<212>dna<213>人工序列<220><223><400>168ggugaggaggaagucuucuaccguuuuagagcuagaaauag41<210>169<211>41<212>dna<213>人工序列<220><223><400>169ggacuccugaaaugauaaaucaguuuuagagcuagaaauag41<210>170<211>41<212>dna<213>人工序列<220><223><400>170ggacuggccagugauccucaugguuuuagagcuagaaauag41<210>171<211>41<212>dna<213>人工序列<220><223><400>171gggagauacugaagauguuccuguuuuagagcuagaaauag41<210>172<211>41<212>dna<213>人工序列<220><223><400>172ggagacagaaugaauguagaaaguuuuagagcuagaaauag41<210>173<211>41<212>dna<213>人工序列<220><223><400>173ggugcuugugaauuuucugagaguuuuagagcuagaaauag41<210>174<211>41<212>dna<213>人工序列<220><223><400>174ggcuucaguccuucuacuguccguuuuagagcuagaaauag41<210>175<211>39<212>dna<213>人工序列<220><223><400>175ggcucuaaucaaucgacuccguuuuagagcuagaaauag39<210>176<211>40<212>dna<213>人工序列<220><223><400>176ggcauguaccugugcuauaugguuuuagagcuagaaauag40<210>177<211>41<212>dna<213>人工序列<220><223><400>177ggagcauucaaagugucaaaguguuuuagagcuagaaauag41<210>178<211>41<212>dna<213>人工序列<220><223><400>178ggcuacaguaggggcauccauaguuuuagagcuagaaauag41<210>179<211>41<212>dna<213>人工序列<220><223><400>179ggacccuacacucuccggaugaguuuuagagcuagaaauag41<210>180<211>41<212>dna<213>人工序列<220><223><400>180ggaucugaggaacccccaucguguuuuagagcuagaaauag41<210>181<211>41<212>dna<213>人工序列<220><223><400>181gguccaauguccagaacacuacguuuuagagcuagaaauag41<210>182<211>41<212>dna<213>人工序列<220><223><400>182ggagugcuagauacuuucacacguuuuagagcuagaaauag41<210>183<211>40<212>dna<213>人工序列<220><223><400>183ggagcagacacgucauauuuaguuuuagagcuagaaauag40<210>184<211>41<212>dna<213>人工序列<220><223><400>184ggaggaucaucaagaauuaugcguuuuagagcuagaaauag41<210>185<211>41<212>dna<213>人工序列<220><223><400>185ggagcgauucacaaaagagcacguuuuagagcuagaaauag41<210>186<211>41<212>dna<213>人工序列<220><223><400>186ggcaccuaacguuuaacaccuaguuuuagagcuagaaauag41<210>187<211>41<212>dna<213>人工序列<220><223><400>187gguuacacucccaagaucaaucguuuuagagcuagaaauag41<210>188<211>41<212>dna<213>人工序列<220><223><400>188ggccaccgugccucgccucaugguuuuagagcuagaaauag41<210>189<211>41<212>dna<213>人工序列<220><223><400>189ggugcacguucuacacguguccguuuuagagcuagaaauag41<210>190<211>40<212>dna<213>人工序列<220><223><400>190gguuguuaagucuuagucauuguuuuagagcuagaaauag40<210>191<211>40<212>dna<213>人工序列<220><223><400>191ggccccagagucagcucguguguuuuagagcuagaaauag40<210>192<211>41<212>dna<213>人工序列<220><223><400>192gguggguacaugaauacaguguguuuuagagcuagaaauag41<210>193<211>41<212>dna<213>人工序列<220><223><400>193gguuagauguacuagucuaucaguuuuagagcuagaaauag41<210>194<211>41<212>dna<213>人工序列<220><223><400>194ggaccaccuaucaucuaaugauguuuuagagcuagaaauag41<210>195<211>40<212>dna<213>人工序列<220><223><400>195ggucaacccugacauauuggcguuuuagagcuagaaauag40<210>196<211>41<212>dna<213>人工序列<220><223><400>196ggcauguuguagcuuauguuauguuuuagagcuagaaauag41<210>197<211>41<212>dna<213>人工序列<220><223><400>197ggcuucuggauucuggcuuauaguuuuagagcuagaaauag41<210>198<211>41<212>dna<213>人工序列<220><223><400>198ggagacagaugcuagcaccaaaguuuuagagcuagaaauag41<210>199<211>41<212>dna<213>人工序列<220><223><400>199ggacaacuaauauaccagucagguuuuagagcuagaaauag41<210>200<211>41<212>dna<213>人工序列<220><223><400>200ggugcuuuguucuggauuucgcguuuuagagcuagaaauag41<210>201<211>39<212>dna<213>人工序列<220><223><400>201ggaauugaaaucaccuaguaguuuuagagcuagaaauag39<210>202<211>41<212>dna<213>人工序列<220><223><400>202ggagguaguaugaguuccaucaguuuuagagcuagaaauag41<210>203<211>41<212>dna<213>人工序列<220><223><400>203gguuuaaggugaagcagcaucuguuuuagagcuagaaauag41<210>204<211>41<212>dna<213>人工序列<220><223><400>204ggauuacugguggacuuacuucguuuuagagcuagaaauag41<210>205<211>41<212>dna<213>人工序列<220><223><400>205ggcucugucaaaugucgugguaguuuuagagcuagaaauag41<210>206<211>41<212>dna<213>人工序列<220><223><400>206gguucuugaucucccacacuauguuuuagagcuagaaauag41<210>207<211>40<212>dna<213>人工序列<220><223><400>207ggaaaugguugcugggcacggguuuuagagcuagaaauag40<210>208<211>41<212>dna<213>人工序列<220><223><400>208gguauagguaaacacacgacuuguuuuagagcuagaaauag41<210>209<211>41<212>dna<213>人工序列<220><223><400>209gguucugaagauaccguuaauaguuuuagagcuagaaauag41<210>210<211>41<212>dna<213>人工序列<220><223><400>210gguuaaaggcaugggcuucgccguuuuagagcuagaaauag41<210>211<211>41<212>dna<213>人工序列<220><223><400>211ggagugaauuuuaugagcccuaguuuuagagcuagaaauag41<210>212<211>40<212>dna<213>人工序列<220><223><400>212ggacgucugucuacauugaauguuuuagagcuagaaauag40<210>213<211>41<212>dna<213>人工序列<220><223><400>213ggcagcuaacauguaugaugccguuuuagagcuagaaauag41<210>214<211>40<212>dna<213>人工序列<220><223><400>214ggccuuagcccccuuaguagcguuuuagagcuagaaauag40<210>215<211>39<212>dna<213>人工序列<220><223><400>215gguuucuguagcccauacuuguuuuagagcuagaaauag39<210>216<211>41<212>dna<213>人工序列<220><223><400>216ggagcauacauaggguuucucuguuuuagagcuagaaauag41<210>217<211>41<212>dna<213>人工序列<220><223><400>217ggcagcacuugagugucauucuguuuuagagcuagaaauag41<210>218<211>41<212>dna<213>人工序列<220><223><400>218ggcaucuguauaaaccgugugaguuuuagagcuagaaauag41<210>219<211>41<212>dna<213>人工序列<220><223><400>219ggauuugguaaugaugcuagguguuuuagagcuagaaauag41<210>220<211>40<212>dna<213>人工序列<220><223><400>220ggccauuuaaaaaguaauggcguuuuagagcuagaaauag40<210>221<211>41<212>dna<213>人工序列<220><223><400>221ggaagcauuggaggaauaucguguuuuagagcuagaaauag41<210>222<211>41<212>dna<213>人工序列<220><223><400>222ggacaacauaaucaucguuugcguuuuagagcuagaaauag41<210>223<211>40<212>dna<213>人工序列<220><223><400>223ggucacugguuaaaacuaaggguuuuagagcuagaaauag40<210>224<211>41<212>dna<213>人工序列<220><223><400>224ggaguaaugcaauaugguagacguuuuagagcuagaaauag41<210>225<211>41<212>dna<213>人工序列<220><223><400>225ggcagugagaauguauauacucguuuuagagcuagaaauag41<210>226<211>41<212>dna<213>人工序列<220><223><400>226gguaauauuuuggcuaagagccguuuuagagcuagaaauag41<210>227<211>41<212>dna<213>人工序列<220><223><400>227gguucucauucauauaaauuguguuuuagagcuagaaauag41<210>228<211>41<212>dna<213>人工序列<220><223><400>228gguuuuacccccagugguauguguuuuagagcuagaaauag41<210>229<211>41<212>dna<213>人工序列<220><223><400>229gguagaaaugcccugaucauuaguuuuagagcuagaaauag41<210>230<211>41<212>dna<213>人工序列<220><223><400>230ggcuaugagcacaguagaacuaguuuuagagcuagaaauag41<210>231<211>41<212>dna<213>人工序列<220><223><400>231gguacauuuagugguaguccagguuuuagagcuagaaauag41<210>232<211>41<212>dna<213>人工序列<220><223><400>232ggcuuaaguauaguaauuagaaguuuuagagcuagaaauag41<210>233<211>40<212>dna<213>人工序列<220><223><400>233ggcuuagauaaauuacagauuguuuuagagcuagaaauag40<210>234<211>41<212>dna<213>人工序列<220><223><400>234gguaguaguccccccuuauccaguuuuagagcuagaaauag41<210>235<211>41<212>dna<213>人工序列<220><223><400>235gguugcgguaaaccgagaucacguuuuagagcuagaaauag41<210>236<211>41<212>dna<213>人工序列<220><223><400>236ggucaggcuuuacuagaagaacguuuuagagcuagaaauag41<210>237<211>41<212>dna<213>人工序列<220><223><400>237ggaauagcugcaaagaccacauguuuuagagcuagaaauag41<210>238<211>41<212>dna<213>人工序列<220><223><400>238ggauucaucagcguuugcuucaguuuuagagcuagaaauag41<210>239<211>41<212>dna<213>人工序列<220><223><400>239ggcaaggaaguuguaccgucuuguuuuagagcuagaaauag41<210>240<211>40<212>dna<213>人工序列<220><223><400>240ggcaguaaagcaggcaauaucguuuuagagcuagaaauag40<210>241<211>41<212>dna<213>人工序列<220><223><400>241ggucuguagcuuugaagaaugcguuuuagagcuagaaauag41<210>242<211>41<212>dna<213>人工序列<220><223><400>242ggacauuugcaaaugcugauucguuuuagagcuagaaauag41<210>243<211>41<212>dna<213>人工序列<220><223><400>243ggagugaccugauucuaaacacguuuuagagcuagaaauag41<210>244<211>41<212>dna<213>人工序列<220><223><400>244ggccaggaguucgagacuagccguuuuagagcuagaaauag41<210>245<211>41<212>dna<213>人工序列<220><223><400>245ggugucccaaaagagcuaguuaguuuuagagcuagaaauag41<210>246<211>41<212>dna<213>人工序列<220><223><400>246gguccacuuuugaauguuguacguuuuagagcuagaaauag41<210>247<211>41<212>dna<213>人工序列<220><223><400>247ggaaaaugucagacaagcucaaguuuuagagcuagaaauag41<210>248<211>41<212>dna<213>人工序列<220><223><400>248ggucuuccaaguagcuaaugaaguuuuagagcuagaaauag41<210>249<211>40<212>dna<213>人工序列<220><223><400>249ggauuacaugaacaaaugggcguuuuagagcuagaaauag40<210>250<211>40<212>dna<213>人工序列<220><223><400>250ggauacaguauuaauugacugguuuuagagcuagaaauag40<210>251<211>41<212>dna<213>人工序列<220><223><400>251gguggucuuuaagauagucaucguuuuagagcuagaaauag41<210>252<211>41<212>dna<213>人工序列<220><223><400>252ggaggggcuuuuauucugcucaguuuuagagcuagaaauag41<210>253<211>41<212>dna<213>人工序列<220><223><400>253ggaauaauauugaaaugacuacguuuuagagcuagaaauag41<210>254<211>41<212>dna<213>人工序列<220><223><400>254ggauuaucuggccaguuuaugaguuuuagagcuagaaauag41<210>255<211>40<212>dna<213>人工序列<220><223><400>255ggaaaaguuaugcaauucuucguuuuagagcuagaaauag40<210>256<211>41<212>dna<213>人工序列<220><223><400>256gguggguuuucauacagcuagcguuuuagagcuagaaauag41<210>257<211>41<212>dna<213>人工序列<220><223><400>257gguuaucacuuaagagcuuaggguuuuagagcuagaaauag41<210>258<211>41<212>dna<213>人工序列<220><223><400>258ggacaacuuguguaaaaagcuaguuuuagagcuagaaauag41<210>259<211>41<212>dna<213>人工序列<220><223><400>259ggauauaccucaucagaaugguguuuuagagcuagaaauag41<210>260<211>41<212>dna<213>人工序列<220><223><400>260ggccugcauuuaggauagccagguuuuagagcuagaaauag41<210>261<211>41<212>dna<213>人工序列<220><223><400>261ggauguagcacgcauucacauaguuuuagagcuagaaauag41<210>262<211>39<212>dna<213>人工序列<220><223><400>262gggauuuuuagcacagcaagguuuuagagcuagaaauag39<210>263<211>40<212>dna<213>人工序列<220><223><400>263gguaaugaagcaucugauaccguuuuagagcuagaaauag40<210>264<211>41<212>dna<213>人工序列<220><223><400>264ggcaaaguuaagggaguguuagguuuuagagcuagaaauag41<210>265<211>40<212>dna<213>人工序列<220><223><400>265ggacaaacaacaguugguauuguuuuagagcuagaaauag40<210>266<211>41<212>dna<213>人工序列<220><223><400>266ggagagaauguguggcaugacuguuuuagagcuagaaauag41<210>267<211>41<212>dna<213>人工序列<220><223><400>267ggcguauacagauuugauaucuguuuuagagcuagaaauag41<210>268<211>41<212>dna<213>人工序列<220><223><400>268ggacuuuacucuuucaaacauuguuuuagagcuagaaauag41<210>269<211>41<212>dna<213>人工序列<220><223><400>269ggcuguucucccucuauagguaguuuuagagcuagaaauag41<210>270<211>41<212>dna<213>人工序列<220><223><400>270ggugcuuguacugugaguuauuguuuuagagcuagaaauag41<210>271<211>41<212>dna<213>人工序列<220><223><400>271gguuuaaacaugucuuaccgaaguuuuagagcuagaaauag41<210>272<211>40<212>dna<213>人工序列<220><223><400>272ggaugaaguacuaagguugagguuuuagagcuagaaauag40<210>273<211>40<212>dna<213>人工序列<220><223><400>273ggcagacccucauuugcuacaguuuuagagcuagaaauag40<210>274<211>41<212>dna<213>人工序列<220><223><400>274ggaugagacacuugauuacuacguuuuagagcuagaaauag41<210>275<211>41<212>dna<213>人工序列<220><223><400>275ggaguuacagcuacugcuugauguuuuagagcuagaaauag41<210>276<211>41<212>dna<213>人工序列<220><223><400>276ggcaaucuaggacugcuguuacguuuuagagcuagaaauag41<210>277<211>41<212>dna<213>人工序列<220><223><400>277ggugaggcgggcagaucaucugguuuuagagcuagaaauag41<210>278<211>41<212>dna<213>人工序列<220><223><400>278ggcucugcguguucucauaaacguuuuagagcuagaaauag41<210>279<211>41<212>dna<213>人工序列<220><223><400>279ggccccuugcuaggccugccucguuuuagagcuagaaauag41<210>280<211>41<212>dna<213>人工序列<220><223><400>280gguagcaggaggcguauaaacgguuuuagagcuagaaauag41<210>281<211>41<212>dna<213>人工序列<220><223><400>281gguuuguguagcuguauacguaguuuuagagcuagaaauag41<210>282<211>41<212>dna<213>人工序列<220><223><400>282ggugucucucgaacuaaaaaguguuuuagagcuagaaauag41<210>283<211>40<212>dna<213>人工序列<220><223><400>283ggaauucaguaucauccuaugguuuuagagcuagaaauag40<210>284<211>41<212>dna<213>人工序列<220><223><400>284ggcguaaucauauacggcaguaguuuuagagcuagaaauag41<210>285<211>41<212>dna<213>人工序列<220><223><400>285gguauuagcaauccccaaauagguuuuagagcuagaaauag41<210>286<211>41<212>dna<213>人工序列<220><223><400>286ggaucggcuauaaaaaagauaaguuuuagagcuagaaauag41<210>287<211>41<212>dna<213>人工序列<220><223><400>287gguggggcuucaagagguguacguuuuagagcuagaaauag41<210>288<211>41<212>dna<213>人工序列<220><223><400>288gguuaagcucaagaaagaucucguuuuagagcuagaaauag41<210>289<211>40<212>dna<213>人工序列<220><223><400>289ggcuuccucaaaaaaucuacaguuuuagagcuagaaauag40<210>290<211>41<212>dna<213>人工序列<220><223><400>290ggaguuugguauaccagcgagcguuuuagagcuagaaauag41<210>291<211>40<212>dna<213>人工序列<220><223><400>291ggagaaucacuugaacccgggguuuuagagcuagaaauag40<210>292<211>41<212>dna<213>人工序列<220><223><400>292ggcaaguuaauuguaucaggccguuuuagagcuagaaauag41<210>293<211>41<212>dna<213>人工序列<220><223><400>293ggagccuuauucacuaaaauucguuuuagagcuagaaauag41<210>294<211>40<212>dna<213>人工序列<220><223><400>294gguaucuaggguauucuuuuuguuuuagagcuagaaauag40<210>295<211>41<212>dna<213>人工序列<220><223><400>295ggucucacucugucacccaaucguuuuagagcuagaaauag41<210>296<211>41<212>dna<213>人工序列<220><223><400>296ggugucuuguuagugcacgugaguuuuagagcuagaaauag41<210>297<211>41<212>dna<213>人工序列<220><223><400>297gguucaacuauauaccgaguagguuuuagagcuagaaauag41<210>298<211>41<212>dna<213>人工序列<220><223><400>298gguagagcucaucuuaagggugguuuuagagcuagaaauag41<210>299<211>41<212>dna<213>人工序列<220><223><400>299ggcaagggaugucacaaccgugguuuuagagcuagaaauag41<210>300<211>41<212>dna<213>人工序列<220><223><400>300ggaaacaguuauacugaguauuguuuuagagcuagaaauag41<210>301<211>41<212>dna<213>人工序列<220><223><400>301ggagugaaggggcucccguggcguuuuagagcuagaaauag41<210>302<211>40<212>dna<213>人工序列<220><223><400>302ggauuaguuggagcuaccaguguuuuagagcuagaaauag40<210>303<211>40<212>dna<213>人工序列<220><223><400>303ggcugaauuuggcgaaagcucguuuuagagcuagaaauag40<210>304<211>41<212>dna<213>人工序列<220><223><400>304ggauuuggauucuggucgccacguuuuagagcuagaaauag41<210>305<211>41<212>dna<213>人工序列<220><223><400>305ggccccaucuccugagguucauguuuuagagcuagaaauag41<210>306<211>40<212>dna<213>人工序列<220><223><400>306ggcaucggcauguuugacaauguuuuagagcuagaaauag40<210>307<211>41<212>dna<213>人工序列<220><223><400>307gguuaguugggguggaccacuuguuuuagagcuagaaauag41<210>308<211>40<212>dna<213>人工序列<220><223><400>308ggcucaaaucauuccugguacguuuuagagcuagaaauag40<210>309<211>41<212>dna<213>人工序列<220><223><400>309ggugaagcaagauaugaaacucguuuuagagcuagaaauag41<210>310<211>41<212>dna<213>人工序列<220><223><400>310ggccuaaccuauuaggaguuagguuuuagagcuagaaauag41<210>311<211>41<212>dna<213>人工序列<220><223><400>311ggcagucuacucaagaaauccaguuuuagagcuagaaauag41<210>312<211>40<212>dna<213>人工序列<220><223><400>312ggcucuuuugucugguucaacguuuuagagcuagaaauag40<210>313<211>40<212>dna<213>人工序列<220><223><400>313ggcaacauaaguacuaaugugguuuuagagcuagaaauag40<210>314<211>30<212>dna<213>人工序列<220><223><400>314acgttggatgacctatcggaagaaggcaag30<210>315<211>30<212>dna<213>人工序列<220><223><400>315acgttggatgcgctcttgtattatctgtgg30<210>316<211>30<212>dna<213>人工序列<220><223><400>316acgttggatgtaacttcagctctgggaaag30<210>317<211>30<212>dna<213>人工序列<220><223><400>317acgttggatgctggagccaagaagagtaac30<210>318<211>30<212>dna<213>人工序列<220><223><400>318acgttggatgagagtgggcagagaatgttg30<210>319<211>30<212>dna<213>人工序列<220><223><400>319acgttggatgaaggtttcaaagcgccagtc30<210>320<211>30<212>dna<213>人工序列<220><223><400>320acgttggatgtgtctaagaacacagaggag30<210>321<211>30<212>dna<213>人工序列<220><223><400>321acgttggatgaggtgatgttcctgagatgc30<210>322<211>31<212>dna<213>人工序列<220><223><400>322acgttggatgctcattctgaatagaatcacc31<210>323<211>30<212>dna<213>人工序列<220><223><400>323acgttggatgaacggagcagaatggtcaag30<210>324<211>30<212>dna<213>人工序列<220><223><400>324acgttggatgacggctaattgtgctcactg30<210>325<211>30<212>dna<213>人工序列<220><223><400>325acgttggatggctagaggaaaactttgagg30<210>326<211>30<212>dna<213>人工序列<220><223><400>326acgttggatgctctgggtccttaaagaaac30<210>327<211>30<212>dna<213>人工序列<220><223><400>327acgttggatgacaggcttgatattagactc30<210>328<211>30<212>dna<213>人工序列<220><223><400>328acgttggatgagatctgaatgctgatcccc30<210>329<211>30<212>dna<213>人工序列<220><223><400>329acgttggatgctaggattctctgagcatgg30<210>330<211>30<212>dna<213>人工序列<220><223><400>330acgttggatgacagtgagcacaattagccg30<210>331<211>30<212>dna<213>人工序列<220><223><400>331acgttggatgttagtactggaacctacttc30<210>332<211>30<212>dna<213>人工序列<220><223><400>332acgttggatgttagtactggaacctacttc30<210>333<211>30<212>dna<213>人工序列<220><223><400>333acgttggatgacagtgagcacaattagccg30當前第1頁12