Mouse Uroplakin II Promotersun-lab.med.nyu.edu/files_private/sun-lab... · Mouse Uroplakin II...

MouseUroplakinIIPromoterTung‐TienSunandXue‐RuWu,NYUSchoolofMedicine

(UpdatedJuly2010)A. Generalinformation:Wehaveisolatedandcharacterizeda3.6kb5’‐upstreamsequenceofmouseuroplakinIIgene,andweshowedthatthissequencecanserveasahighlybladderurothelium‐specificpromoterintransgenicmousestudies.Thissequencehasbeenusedtodrivethemousebladder‐specificexpressionofhumangrowthfactor,SV40‐largeTantigen,Ha‐Ras,EGFreceptor.Ithasalsobeenusedtoachievetheurothelium‐specificablation(conventionalorinducible)ofvariousgenesincludingRbandp53.ThesestudieshaveenabledDr.Xue‐RuWutodissecttherolesofvariousoncogenesandtumore‐suppressorgenesinbladdertumorigenesis.ThispagecontainsthesequenceandrestrictionmapofthismouseUPIIpromoter(updatedfromtheoriginalpaperbyLinetalPNAS1995).Blastingthissequenceagainstthemousegenome(NCBI)showed99%identity.Whiletheseminordifferencesmaynotaffecttheutilityofthispromoterinyourtransgenicexperiments,youshouldkeepsuchdifferencesinmindifyouseeanunexpectedrestrictionenzymecutpattern.ThispromoterisinapBluescriptSK(+)vector,whichcontainstheAmp‐resistantgene.Toamplify,transformthisintoJM109orDH5alphaandselectonAmpplatesandbroth.WerecommendtheuseofKpnIasa5’restrictionenzymetocutoutthepromoter.TheXhoIandApaIenzymesdonotcutduetoadisruptionduringthesubcloningprocess.Usethemaptofigureoutwhatenzymeyoupreferforthe3'cut.VisitStratagenforadditionalinformationaboutthevectorsequenceandrestrictionmap.Ifyouliketousethispromoterforyourresearchorhaveanyquestions,pleasecontactDr.Tung‐TienSunatsunt01@nyumc.orgorDr.Xue‐RuWuatXue‐Ru.Wu@nyumc.orgB. References:Lin,J.H.,Zhao,H.,andSun,T.‐T.(1995).Atissue‐specificpromoterthatcandriveaforeigngenetoexpressinthesuprabasalurothelialcellsoftransgenicmice.ProcNatlAcadSci(USA)92,679‐683.Kerr,D.E.,Liang,F.,Bondioli,K.R.,Zhao,H.,Kreibich,G.,Wall,R.J.,andSun,T.‐T.(1998).Thebladderasabioreactor:urotheliumproductionandsecretionofgrowthhormoneintourine.NatureBiotechnol16,75‐79.

Zhang,Z.T.,Pak,J.,Shapiro,E.,Sun,T.‐T.,andWu,X.R.(1999).Urothelium‐specificexpressionofanoncogeneintransgenicmiceinducedtheformationofcarcinomainsituandinvasivetransitionalcellcarcinoma.CancerRes59,3512‐3517.Zhang,Z.T.,Pak,J.,Huang,H.Y.,Shapiro,E.,Sun,T.T.,Pellicer,A.,andWu,X.R.(2001).RoleofHa‐rasactivationinsuperficialpapillarypathwayofurothelialtumorformation.Oncogene20,1973‐1980.Cheng,J.,Huang,H.,Zhang,Z.T.,Shapiro,E.,Pellicer,A.,Sun,T.T.,andWu,X.R.(2002).Overexpressionofepidermalgrowthfactorreceptorinurotheliumelicitsurothelialhyperplasiaandpromotesbladdertumorgrowth.CancerRes62,4157‐4163.Cheng,J.,Huang,H.,Pak,J.,Shapiro,E.,Sun,T.T.,Cordon‐Cardo,C.,Waldman,F.M.,andWu,X.‐R.(2003).Alleliclossofp53geneisassociatedwithmaintenance,butnotprogression,ofmousecarcinomainsituofthebladder.CancerRes63,179‐185.Gao,J.,Huang,H.Y.,Pak,J.,Cheng,J.,Zhang,Z.T.,Shapiro,E.,Pellicer,A.,Sun,T.T.,andWu,X.R.(2004).p53deficiencyprovokesurothelialproliferationandsynergizeswithactivatedHa‐rasinpromotingurothelialtumorigenesis.Oncogene23,687‐696.Mo,L.,Cheng,J.,Lee,E.Y.,Sun,T.T.,andWu,X.R.(2005).GenedeletioninurotheliumbyspecificexpressionofCrerecombinase.AmJPhysiolRenalPhysiol289,F562‐568.Salz,W.,Eisenberg,D.,Plescia,J.,Garlick,D.S.,Weiss,R.M.,Wu,X.R.,Sun,T.T.,andAltieri,D.C.(2005).Asurvivingenesignaturepredictsaggressivetumorbehavior.CancerRes65,3531‐3534.Wu,Xue‐Ru."Urothelialtumorigenesis:ataleofdivergentpathways".Naturereviews.Cancer.2005;5:713Johnson,A.M.,Conover,D.L.,Huang,J.,Messing,E.M.,Ning,R.,O'Connell,M.J.,Rossi,M.A.,Sun,T.T.,Wood,R.W.,Wu,X.R.,etal.(2006).Earlydetectionandmeasurementofurothelialtumorsinmice.Urology67,1309‐1314.Liu,W.,Evanoff,D.P.,Chen,X.,andLuo,Y.(2007).UrinarybladderepitheliumantigeninducesCD8+Tcelltolerance,activation,andautoimmuneresponse.JImmunol178,539‐546.Liu,W.,Deyoung,B.R.,Chen,X.,Evanoff,D.P.,andLuo,Y.(2008).RDP58inhibitsTcell‐mediatedbladderinflammationinanautoimmunecystitismodel.JAutoimmun30,257‐265.

Wu,Xue‐Ru."Biologyofurothelialtumorigenesis:insightsfromgeneticallyengineeredmice".CancerMetastasisReviews.2009;28:281Schnegelsberg,B.,Sun,T.T.,Cain,G.,Bhattacharya,A.,Nunn,P.A.,Ford,A.P.,Vizzard,M.A.,andCockayne,D.A.(2010).OverexpressionofNGFinmouseurotheliumleadstoneuronalhyperinnervation,pelvicsensitivity,andchangesinurinarybladderfunction.AmJPhysiolRegulIntegrCompPhysiol298,R534‐547.Zhou,H.,Liu,Y.,He,F.,Mo,L.,Sun,T.T.,andWu,X.R.(2010).TemporallyandSpatiallyControllableGeneExpressionandKnockoutinMouseUrothelium.AmJPhysiolRenalPhysiol.C. Sequence:>NAME=up2:TYPE=DNACTCGAGGATCTCGGCCCTCTTTCTGCATCCTTGTCCTAAATCATTTTCATATCTTGCTAGACCTCAGTTTGAGAGAAACGAACCTTCTCATTTTCAAGTTGAAAAAAAAAAGAGGTTCAAAGTGGCTCACTCAAAGTTACAAGCCAACACTCACCACTACGAGTACAATGGCCACCATTAGTGTCTGGCATGCCCCAGGAGACAGGCATGCATATTATTCTAGATGACTGGGAGGCAGAGGGGTGGCCTAGTGAGGTCAGACTGTGGACAGATCAGGCAGATGTGGGTTCTGATCCCAATTCCTCAGGCCGCAGAACTACTGTGGTTCAAGAAGGGGACAAAAGGACTGCAGTCCCGGAACAGGAGGTCCACTTTGAGAGCTGACTGAGCAGAAGAGGAAAGTGAAGAACTTCAGGGGCAAGAGCTTACCCTACTTTTACAGCTTTGTTGTCTTCTTTACTCCAGGGGCGTCCCTGGTACTCAGTAAATGTCTGTTGGCTTGAGGAACATATGTGTAAGGAGGAAGGAGAGGGAACTTGAGGGAGTTAAGACTCAAGAATCAATCAAGGAGAGGACAGCAGAGAAGACAGGGTTTGGGAGAGAGACTCCAGACATTGGCCCTGGTTCCCTTCTTGGCCACTGTGAAACCCTCCAGAGGAACTGAGTGCTGTGGCTTTAAATGATCTCAGCACTGTCAGTGAAGCCGCTCTGCTCAAAGAGTTATCCTCTTGCTCCTGTGCCGGGGCCTCCCCCTCCTCTCAGCTCCCCAAACCCTTCTCAGCCACTGTGATGGGATAATTAGATGCGAGAGCTCAGCACAGATGATGCTCCAGTTGCTTAGCAACTAATGGTTTCCATGGAGACCGCAAAGCACAGCCTCTAGAGCAGCCAGTGAGCAGCTCGGCAGGGCAGGGAGAAGACGCAACTCTGCTCCTCCAGAAACCTGGGGAGGGTTAGGGTGGGGAAGGTGAGGGGGGATTGGAGGGAGATGGGAAGGCTTAAAGGCACAGGACCCTCATATCCCTTTAAAAATGTGACCAGAGATCTGCTCTTTTTATCCCACACTGTCTCACTTCTAATTTCAAATGGGGCTTAGGAGTGGACCTATATGGTCTGCTCTGCAGTAAACCAATCCTCTTCCTGATTCCTCCTAAATCCTCACGTAAACACTGTGCTATGTGGAGGTTTGAGACTTGCCAGGAAGATCTCCTGACCCTGACACCCTACCGTGTAGCCCTCTTCCGCACCGCCAGTTCTCCCTCGTGTGTGATCTTTCTGTGTGTAAAGTGAGGAGGAGATCTCATCTAAAGGGGACAAAAGCCAAGGAGCTGGAAAAACAGAAATCATTCAGTTCTCCACCCAAAGGCAGCCTGAGAAAAACCTCTGTCCCACTTTGCATTTGCATGTGTGTGTGTGTATGAGTGTGTGTGTGTATGAGTGTGTGTGTGTATGAGTGTGTGTGTGTATGAGTGTGTGTGTGTATGAGTGTGTGTGTGTGGATGAGTGTGTGTGTGCACATGTATGAGTGTGTCTGTGTATGTATGATTATATGTGTGTGTCTGTGTGTGTGTATGAGTATGTGTGTCTGTGTGTGTGTATGAGTATGTGTGTCTGTGTGTGTGGATGAGTATGTGTGTCTGTGTGTGTGGATGAGTGTGTGTGTGGATGTGTATGAATATGTGAATGTGAGAGTGCATGTGTGTGTGTCTGTGTGTATACGAGTATGTGTGTGCATGTGGATAAGTGTGTGTGTGTGTGTATGAGTATGTGTGTGTGTCTGTGTATGAGTGTTTACACCAATACAGCATCTGCTAACTCCAAATGTGAGTCTGTGTCTCTGCGTTACCTGTAAGCGTCATAGGAAGTTTTACGAAACCATGAGATGGCTCAACAGGTAAAAGCTCTTGCCACCATGCCTGACATTTTGGTCCCCAGAAGCCACATTATGAAAGGAGAGAGCCAATGCCCACAACTTGCCCTCTGACCTCCACATAGACACTGTGGCACACATATGGGCACACATATAAAATAATAAATGCAACTTTAAATTTTTGAAAGAAATTAAGTTTCGGGTTTTACCTCAGACCCACTGTATCAGAAGCTACATCTGAACCAGTTTCCTGATCGTCCATTGCACAGTAAATTTGGAGATCGTCTGGTGAGAGTTCCTCCTGCTCCCAGCCGGAGCCCCCAGGTTCTGTTGTCTTCTACCCCCTACACACATCGCAGACTTTAAGTCCACCCAGCATGGGGGAAAGGACAGAACCCCATCAGCCCTTCAGTGGCCCCCACTTTTCCACTAGGACCTTTCTCTGGGGTTGGGGAAAATGGGTGGTTCCTGGAGCAGACCCTGACGGTCTGAGCTCAGGTCCTATCGAGTTCACCTAGCTGAGACACCCACACCCCTGCAGCCACTTTGCAGTGACAACCTGAGTCTCAGGTTCTGCATCTATAAAAAGCAGTGCCTTTCAGGAGGGCATGCAGAGCCCCCTGGCCAGCGTCTAGAGGAGAGGTGACTGAGTGGGGCCATGTCACTCGTCCATGGCTGGAGAACCTCCATCAGTTTCTCCCAGTTAGCCTGGGGCAGGAGAGAACCAGAGGAGCTGTGGCTGCTGATGGTGTTTCTTCCCCAACTGTTGTCCCAGGCATCGAACCCCAGAGCGACCTGCACACAGTCCACCGCTGAGCCACACCCCAGCCCTCCACCTCCTCTGCTCCTGGTTACAGGATTGTTTTGTCTTGAAGGGTTTTGTTGTTGCTACTTTTTGCTTTGTTTTTTCTTTTTTCTTTTTTAACATAAGGTTTCTCTGTGTAGCCCTAGCTGTCCTGGAACTCACTCTGTAGACCAGGCTGGCCTCAAACTCAGAAATCCACCTTCCTCCCAAGTGCTGGGATTAAAGGCATTCGCCACCATCGCCCAGCCCCCGGTCTTGTTTCCTAAGGTTTTCCTGCTTTACTCGCTACCCGTTGCACAACCGCTTGCTGTCCAAGTCTGTTTGTATCTACTCCACCGCCCACTAGCCTTGCTGGACTGGACCTACGTTTACCTGGAAGCCTTCACTAACTTCCCTTGTCTCCACCTTCTGGAGAAATCTGAAGGCTCACACTGATACCCTCCGCTTCTCCCAGAGTCGCAGTTTCTTAGGCCTCAGTTAAATACCAGAATTGGATCTCAGGCTCTGCTATCCCCAC

CCTACCTAACCAACCCCCTCCTCTCCCATCCTTACTAGCCAAAGCCCTTTCAACCCTTGGGGCTTTTCCTACACCTACACACCAGGGCAATTTTAGAACTCATGGCTCTCCTAGGAAAACGCCTACCTCCTTGGAGACAGCACCCTCTACAGTCCAGGAGGCAGACACTCAGACAGAGGAACTCTGTCCTTCAGTCGCGGGAGTTCCAGAAAGAGCCATACTCCCCTGCAGCTAACTAAGCTGCCAGGACCCAGCCAGAGCATCCCCCTTTAGGCCCGAGGGCCAGCTCCCCAGCCTGAAAAACCTGTCTGGGGCCCCTCCCTGAGGCTACAGTGCCCAAGGGGCAAGTTGGACTGGATTCCCAGCAGCCCCTCCCACTCCGAGACAAAATCAGCTACCCTGGGGCAGGGCCTCATTGGCCCCAGGAAACCCCAGCCTGTCAGCACCTGTTCCAGG

D. Restrictionenzymesthatdonotcutthepromotersequence(Promega):AatIIAcc65IAccIIIAgeIBamHIBanIBclIBsaOIBsrBRIBssHIIBst98IBstEIIBstZICfoIClaICsp45ICspIEco47IIIEco52IEcoRIEcoRVHaeIIHhaIHincIIHindIIIHpaII‐PpoIKpnIMluINaeINarINgoMIVNheINotINruIPvuIPvuIISacIISalIScaISgfISmaISnaBISpeISspITth111IVspIXmaI

E. Restrictionmaps:SequenceRange:1to3648>XhoII|>Sau3AI|>MboI|>NdeII>HaeIII||>TaqI|>Sau96I||||>AvaI|>FokI||||||>XhoI>DpnI||||||||||||||10|||20304050CTCGAGGATCTCGGCCCTCTTTCTGCATCCTTGTCCTAAATCATTTTCATGAGCTCCTAGAGCCGGGAGAAAGACGTAGGAACAGGATTTAGTAAAAGTA>DdeI|60|708090100ATCTTGCTAGACCTCAGTTTGAGAGAAACGAACCTTCTCATTTTCAAGTTTAGAACGATCTGGAGTCAAACTCTCTTTGCTTGGAAGAGTAAAAGTTCAA110120130140150GAAAAAAAAAAGAGGTTCAAAGTGGCTCACTCAAAGTTACAAGCCAACACCTTTTTTTTTTCTCCAAGTTTCACCGAGTGAGTTTCAATGTTCGGTTGTG>BstOI|>Alw26I||>Hsp92II|||>BalI>BbuI|||||>RsaI>HaeIII>SphI||||||160|170|180190|||200TCACCACTACGAGTACAATGGCCACCATTAGTGTCTGGCATGCCCCAGGAAGTGGTGATGCTCATGTTACCGGTGGTAATCACAGACCGTACGGGGTCCT>NsiI|>BbuI||>SphI||>Hsp92II>XbaI>BsrSI>HaeIII|||||

210|220230|240|250GACAGGCATGCATATTATTCTAGATGACTGGGAGGCAGAGGGGTGGCCTACTGTCCGTACGTATAATAAGATCTACTGACCCTCCGTCTCCCCACCGGAT>DpnI>DpnI||>MboI>NdeII||||>Sau3AI>Sau3AI||||>NdeII>MboI||||260270|280290||300GTGAGGTCAGACTGTGGACAGATCAGGCAGATGTGGGTTCTGATCCCAATCACTCCAGTCTGACACCTGTCTAGTCCGTCTACACCCAAGACTAGGGTTA>HaeIII|>DdeI|||>Bsu36I||||310320330340350TCCTCAGGCCGCAGAACTACTGTGGTTCAAGAAGGGGACAAAAGGACTGCAGGAGTCCGGCGTCTTGATGACACCAAGTTCTTCCCCTGTTTTCCTGACG>HpaII|>NciI>SinI||>MspI>Sau96I>DdeI|||>PstI|>AvaII>AluI||||||||360|370380|390400AGTCCCGGAACAGGAGGTCCACTTTGAGAGCTGACTGAGCAGAAGAGGAATCAGGGCCTTGTCCTCCAGGTGAAACTCTCGACTGACTCGTCTTCTCCTT>XmnI>MboII||>MboII|>MboII>AluI>AluI|||||||410|420|430440||450AGTGAAGAACTTCAGGGGCAAGAGCTTACCCTACTTTTACAGCTTTGTTGTCACTTCTTGAAGTCCCCGTTCTCGAATGGGATGAAAATGTCGAAACAAC>Hsp92I>RsaI||>BstOI|>BstOI|>DdeI|||||460|470|480490500TCTTCTTTACTCCAGGGGCGTCCCTGGTACTCAGTAAATGTCTGTTGGCTAGAAGAAATGAGGTCCCCGCAGGGACCATGAGTCATTTACAGACAACCGA>HinfI|>NdeI>Tru9I|

|||510520530540|550TGAGGAACATATGTGTAAGGAGGAAGGAGAGGGAACTTGAGGGAGTTAAGACTCCTTGTATACACATTCCTCCTTCCTCTCCCTTGAACTCCCTCAATTC>Alw26I|>HinfI>MboII||||560570580590||600ACTCAAGAATCAATCAAGGAGAGGACAGCAGAGAAGACAGGGTTTGGGAGTGAGTTCTTAGTTAGTTCCTCTCCTGTCGTCTCTTCTGTCCCAAACCCTC>HaeIII|>Sau96I>BalI|||>HinfI>AccB7I||>BstOI>HaeIII|||||||610||620|630|640650AGAGACTCCAGACATTGGCCCTGGTTCCCTTCTTGGCCACTGTGAAACCCTCTCTGAGGTCTGTAACCGGGACCAAGGGAAGAACCGGTGACACTTTGGG>NdeII|>MboI|>Sau3AI|>DraI|>DdeI|||>DdeI>Tru9I>DpnI||||||660|670||680|||690700TCCAGAGGAACTGAGTGCTGTGGCTTTAAATGATCTCAGCACTGTCAGTGAGGTCTCCTTGACTCACGACACCGAAATTTACTAGAGTCGTGACAGTCAC>HaeIII|>Sau96I||>NciI||||>MspI||||||>HpaII|||||710720730740|||750AAGCCGCTCTGCTCAAAGAGTTATCCTCTTGCTCCTGTGCCGGGGCCTCCTTCGGCGAGACGAGTTTCTCAATAGGAGAACGAGGACACGGCCCCGGAGG>AluI|>DdeI|>DdeI>BstXI||||760|770|780790800CCCTCCTCTCAGCTCCCCAAACCCTTCTCAGCCACTGTGATGGGATAATT

GGGAGGAGAGTCGAGGGGTTTGGGAAGAGTCGGTGACACTACCCTATTAA>Bsp1286I|>BanII|>SacI|>DdeI||>AluI|||>EcoICRI>BsrSI>DdeI|||||810|||820830|840850AGATGCGAGAGCTCAGCACAGATGATGCTCCAGTTGCTTAGCAACTAATGTCTACGCTCTCGAGTCGTGTCTACTACGAGGTCAACGAATCGTTGATTAC>Hsp92II|>StyI|||>NcoI|>AluI|||>Alw26I|>XbaI>BsrSI>Bst71I|||||||||860870880890|900GTTTCCATGGAGACCGCAAAGCACAGCCTCTAGAGCAGCCAGTGAGCAGCCAAAGGTACCTCTGGCGTTTCGTGTCGGAGATCTCGTCGGTCACTCGTCG>BstOI|>Bst71I>MboII>BstXI||||910920930940||950TCGGCAGGGCAGGGAGAAGACGCAACTCTGCTCCTCCAGAAACCTGGGGAAGCCGTCCCGTCCCTCTTCTGCGTTGAGACGAGGAGGTCTTTGGACCCCT>Tru9I|9609709809901000GGGTTAGGGTGGGGAAGGTGAGGGGGGATTGGAGGGAGATGGGAAGGCTTCCCAATCCCACCCCTTCCACTCCCCCCTAACCTCCCTCTACCCTTCCGAA>DpnI|>BglII||>NdeII||>Sau96I>Sau3AI|||>SinI>DraI>MboI||||>AvaII>Tru9I>XhoII|||||10101020|10301040||1050

AAAGGCACAGGACCCTCATATCCCTTTAAAAATGTGACCAGAGATCTGCTTTTCCGTGTCCTGGGAGTATAGGGAAATTTTTACACTGGTCTCTAGACGA>Alw26I>DdeI||10601070|10801090|1100CTTTTTATCCCACACTGTCTCACTTCTAATTTCAAATGGGGCTTAGGAGTGAAAAATAGGGTGTGACAGAGTGAAGATTAAAGTTTACCCCGAATCCTCA>Sau96I|>SinI>MboII||>AvaII>PstI|>HinfI|||||11101120|11301140|1150GGACCTATATGGTCTGCTCTGCAGTAAACCAATCCTCTTCCTGATTCCTCCCTGGATATACCAGACGAGACGTCATTTGGTTAGGAGAAGGACTAAGGAG>Alw26I>BstOI||116011701180|11901200CTAAATCCTCACGTAAACACTGTGCTATGTGGAGGTTTGAGACTTGCCAGGATTTAGGAGTGCATTTGTGACACGATACACCTCCAAACTCTGAACGGTC>DpnI|>BglII||>MboI||>Sau3AI||>XhoII||>NdeII>MboII>MboII>BsrSI|||||||1210|1220123012401250GAAGATCTCCTGACCCTGACACCCTACCGTGTAGCCCTCTTCCGCACCGCCTTCTAGAGGACTGGGACTGTGGGATGGCACATCGGGAGAAGGCGTGGCG>DpnI|>XhoII||>DpnI>Sau3AI|||>Sau3AI>BglII||||>NdeII>NdeII||||>MboI>MboI||||1260127012801290|1300CAGTTCTCCCTCGTGTGTGATCTTTCTGTGTGTAAAGTGAGGAGGAGATCGTCAAGAGGGAGCACACACTAGAAAGACACACATTTCACTCCTCCTCTAG

>AluI|>StyI>BstXI>XmnI|||13101320|13301340|1350TCATCTAAAGGGGACAAAAGCCAAGGAGCTGGAAAAACAGAAATCATTCAAGTAGATTTCCCCTGTTTTCGGTTCCTCGACCTTTTTGTCTTTAGTAAGT>DdeI>Bst71I||13601370|138013901400GTTCTCCACCCAAAGGCAGCCTGAGAAAAACCTCTGTCCCACTTTGCATTCAAGAGGTGGGTTTCCGTCGGACTCTTTTTGGAGACAGGGTGAAACGTAA>Hsp92II||14101420143014401450TGCATGTGTGTGTGTGTATGAGTGTGTGTGTGTATGAGTGTGTGTGTGTAACGTACACACACACACATACTCACACACACACATACTCACACACACACAT14601470148014901500TGAGTGTGTGTGTGTATGAGTGTGTGTGTGTATGAGTGTGTGTGTGTGGAACTCACACACACACATACTCACACACACACATACTCACACACACACACCT>Bsp1286I|>Alw44I|||>FokI>Hsp92II||||1510|||1520153015401550TGAGTGTGTGTGTGCACATGTATGAGTGTGTCTGTGTATGTATGATTATAACTCACACACACACGTGTACATACTCACACAGACACATACATACTAATAT15601570158015901600TGTGTGTGTCTGTGTGTGTGTATGAGTATGTGTGTCTGTGTGTGTGTATGACACACACAGACACACACACATACTCATACACACAGACACACACACATAC>FokI|161016201630|16401650AGTATGTGTGTCTGTGTGTGTGGATGAGTATGTGTGTCTGTGTGTGTGGATCATACACACAGACACACACACCTACTCATACACACAGACACACACACCT>FokI>FokI>Hsp92II|||1660|1670168016901700TGAGTGTGTGTGTGGATGTGTATGAATATGTGAATGTGAGAGTGCATGTGACTCACACACACACCTACACATACTTATACACTTACACTCTCACGTACAC>AccI>Hsp92II||1710|17201730|17401750TGTGTGTCTGTGTGTATACGAGTATGTGTGTGCATGTGGATAAGTGTGTGACACACAGACACACATATGCTCATACACACACGTACACCTATTCACACAC17601770178017901800

TGTGTGTGTATGAGTATGTGTGTGTGTCTGTGTATGAGTGTTTACACCAAACACACACATACTCATACACACACACAGACACATACTCACAAATGTGGTT>HinfI>Alw26I||18101820|183018401850TACAGCATCTGCTAACTCCAAATGTGAGTCTGTGTCTCTGCGTTACCTGTATGTCGTAGACGATTGAGGTTTACACTCAGACACAGAGACGCAATGGACA>AccB7I|>Hsp92II||186018701880|18901900AAGCGTCATAGGAAGTTTTACGAAACCATGAGATGGCTCAACAGGTAAAATTCGCAGTATCCTTCAAAATGCTTTGGTACTCTACCGAGTTGTCCATTTT>AvaII|>SinI|>AluI>Hsp92II>Sau96I||||1910|1920193019401950GCTCTTGCCACCATGCCTGACATTTTGGTCCCCAGAAGCCACATTATGAACGAGAACGGTGGTACGGACTGTAAAACCAGGGGTCTTCGGTGTAATACTT19601970198019902000AGGAGAGAGCCAATGCCCACAACTTGCCCTCTGACCTCCACATAGACACTTCCTCTCTCGGTTACGGGTGTTGAACGGGAGACTGGAGGTGTATCTGTGA>DraI|>NdeI>Bsp1286I>Tru9I||||2010202020302040||2050GTGGCACACATATGGGCACACATATAAAATAATAAATGCAACTTTAAATTCACCGTGTGTATACCCGTGTGTATATTTTATTATTTACGTTGAAATTTAA>Tru9I>DdeI||2060|2070208020902100TTTGAAAGAAATTAAGTTTCGGGTTTTACCTCAGACCCACTGTATCAGAAAAACTTTCTTTAATTCAAAGCCCAAAATGGAGTCTGGGTGACATAGTCTT>DpnI|>MboI>Sau3AI|||>Sau3AI>MboI|||>AluI>BsrSI>NdeII>NdeII||||||2110|2120||213021402150GCTACATCTGAACCAGTTTCCTGATCGTCCATTGCACAGTAAATTTGGAGCGATGTAGACTTGGTCAAAGGACTAGCAGGTAACGTGTCATTTAAACCTC

>MboII|>BstOI|||>HpaII>BanII|||||>DpnI>MspI>Bsp1286I|||||||216021702180|2190||2200ATCGTCTGGTGAGAGTTCCTCCTGCTCCCAGCCGGAGCCCCCAGGTTCTGTAGCAGACCACTCTCAAGGAGGACGAGGGTCGGCCTCGGGGGTCCAAGAC>EclHKI|>Tru9I>Hsp92II|||221022202230||22402250TTGTCTTCTACCCCCTACACACATCGCAGACTTTAAGTCCACCCAGCATGAACAGAAGATGGGGGATGTGTGTAGCGTCTGAAATTCAGGTGGGTCGTAC>HaeIII|>Sau96I||226022702280||22902300GGGGAAAGGACAGAACCCCATCAGCCCTTCAGTGGCCCCCACTTTTCCACCCCCTTTCCTGTCTTGGGGTAGTCGGGAAGTCACCGGGGGTGAAAAGGTG>Sau96I|>AvaII|>SinI>BstOI|||23102320233023402350TAGGACCTTTCTCTGGGGTTGGGGAAAATGGGTGGTTCCTGGAGCAGACCATCCTGGAAAGAGACCCCAACCCCTTTTACCCACCAAGGACCTCGTCTGG>Sau96I|>AvaII|>SinI|>BanII||>Bsp1286I||>DdeI||||>EcoICRI|>DdeI|||||>AluI|>AluI||||||>DdeI|>SacI>TaqI>Alw26I||||||||||2360|||2370|2380|23902400

CTGACGGTCTGAGCTCAGGTCCTATCGAGTTCACCTAGCTGAGACACCCAGACTGCCAGACTCGAGTCCAGGATAGCTCAAGTGGATCGACTCTGTGGGT>HinfI>Alw26I||>PstI>Bst71I>DdeI|>DdeI||||||241024202430||24402450CACCCCTGCAGCCACTTTGCAGTGACAACCTGAGTCTCAGGTTCTGCATCGTGGGGACGTCGGTGAAACGTCACTGTTGGACTCAGAGTCCAAGACGTAG>BalI|>HaeIII|>SphI>BstOI|||>BbuI>Bsp1286I||||>Hsp92II>BanII|||||246024702480|2490||2500TATAAAAAGCAGTGCCTTTCAGGAGGGCATGCAGAGCCCCCTGGCCAGCGATATTTTTCGTCACGGAAAGTCCTCCCGTACGTCTCGGGGGACCGGTCGC>Hsp92II>Hsp92II||>HaeIII|>StyI|||||>XbaI>DdeI>Sau96I|>NcoI|||||||||25102520||2530|2540|2550TCTAGAGGAGAGGTGACTGAGTGGGGCCATGTCACTCGTCCATGGCTGGAAGATCTCCTCTCCACTGACTCACCCCGGTACAGTGAGCAGGTACCGACCT>BsrSI>BstOI>Bst71I|||2560257025802590|2600GAACCTCCATCAGTTTCTCCCAGTTAGCCTGGGGCAGGAGAGAACCAGAGCTTGGAGGTAGTCAAAGAGGGTCAATCGGACCCCGTCCTCTCTTGGTCTC>TaqI|>AluI>MboII>BstOI||||||2610|262026302640|2650GAGCTGTGGCTGCTGATGGTGTTTCTTCCCCAACTGTTGTCCCAGGCATCCTCGACACCGACGACTACCACAAAGAAGGGGTTGACAACAGGGTCCGTAG>DdeI|>MspA1I||266026702680||26902700GAACCCCAGAGCGACCTGCACACAGTCCACCGCTGAGCCACACCCCAGCCCTTGGGGTCTCGCTGGACGTGTGTCAGGTGGCGACTCGGTGTGGGGTCGG

>BstOI|27102720273027402750CTCCACCTCCTCTGCTCCTGGTTACAGGATTGTTTTGTCTTGAAGGGTTTGAGGTGGAGGAGACGAGGACCAATGTCCTAACAAAACAGAACTTCCCAAA>Tru9I|2760277027802790|2800TGTTGTTGCTACTTTTTGCTTTGTTTTTTCTTTTTTCTTTTTTAACATAAACAACAACGATGAAAAACGAAACAAAAAAGAAAAAAGAAAAAATTGTATT>BstOI|>AluI>BstOI>AccI|||||28102820|28302840|2850GGTTTCTCTGTGTAGCCCTAGCTGTCCTGGAACTCACTCTGTAGACCAGGCCAAAGAGACACATCGGGATCGACAGGACCTTGAGTGAGACATCTGGTCC>HaeIII>DdeI>AccB7I>Tru9I|||||2860|287028802890|2900CTGGCCTCAAACTCAGAAATCCACCTTCCTCCCAAGTGCTGGGATTAAAGGACCGGAGTTTGAGTCTTTAGGTGGAAGGAGGGTTCACGACCCTAATTTC>NciI|>HpaII>Bsu36I||>BsaMI>MspI>DdeI||||29102920|293029402950GCATTCGCCACCATCGCCCAGCCCCCGGTCTTGTTTCCTAAGGTTTTCCTCGTAAGCGGTGGTAGCGGGTCGGGGGCCAGAACAAAGGATTCCAAAAGGA29602970298029903000GCTTTACTCGCTACCCGTTGCACAACCGCTTGCTGTCCAAGTCTGTTTGTCGAAATGAGCGATGGGCAACGTGTTGGCGAACGACAGGTTCAGACAAACA>BsrSI|>Sau96I||>SinI||>AvaII>BstOI|||301030203030||30403050ATCTACTCCACCGCCCACTAGCCTTGCTGGACTGGACCTACGTTTACCTGTAGATGAGGTGGCGGGTGATCGGAACGACCTGACCTGGATGCAAATGGAC>AccB7I|>Alw26I|||306030703080|30903100

GAAGCCTTCACTAACTTCCCTTGTCTCCACCTTCTGGAGAAATCTGAAGGCTTCGGAAGTGATTGAAGGGAACAGAGGTGGAAGACCTCTTTAGACTTCC>HaeIII|>StuI|>HinfI>DdeI>DdeI||||3110312031303140||3150CTCACACTGATACCCTCCGCTTCTCCCAGAGTCGCAGTTTCTTAGGCCTCGAGTGTGACTATGGGAGGCGAAGAGGGTCTCAGCGTCAAAGAATCCGGAG>DdeI|>DpnI||>MboI||||>Sau3AI||||>NdeII||||>Tru9I>XhoII||||||31603170|318031903200AGTTAAATACCAGAATTGGATCTCAGGCTCTGCTATCCCCACCCTACCTATCAATTTATGGTCTTAACCTAGAGTCCGAGACGATAGGGGTGGGATGGAT>FokI>StyI|||32103220323032403250ACCAACCCCCTCCTCTCCCATCCTTACTAGCCAAAGCCCTTTCAACCCTTTGGTTGGGGGAGGAGAGGGTAGGAATGATCGGTTTCGGGAAAGTTGGGAA>BstOI>Hsp92II||32603270|32803290|3300GGGGCTTTTCCTACACCTACACACCAGGGCAATTTTAGAACTCATGGCTCCCCCGAAAAGGATGTGGATGTGTGGTCCCGTTAAAATCTTGAGTACCGAG>StyI|>StyI>Alw26I>BstOI|||||33103320|333033403350TCCTAGGAAAACGCCTACCTCCTTGGAGACAGCACCCTCTACAGTCCAGGAGGATCCTTTTGCGGATGGAGGAACCTCTGTCGTGGGAGATGTCAGGTCC>DdeI|33603370338033903400AGGCAGACACTCAGACAGAGGAACTCTGTCCTTCAGTCGCGGGAGTTCCATCCGTCTGTGAGTCTGTCTCCTTGAGACAGGAAGTCAGCGCCCTCAAGGT>SinI|

>Sau96I|>AluI>AluI>AvaII|||>PstI>Bst71I>FokI||||>Bst71I|>DdeI|>BstOI|||||||34103420||3430|34403450GAAAGAGCCATACTCCCCTGCAGCTAACTAAGCTGCCAGGACCCAGCCAGCTTTCTCGGTATGAGGGGACGTCGATTGATTCGACGGTCCTGGGTCGGTC>AvaI>AluI||>HaeIII>HaeIII|||||>Sau96I|>Sau96I|||||||3460||3470||348034903500AGCATCCCCCTTTAGGCCCGAGGGCCAGCTCCCCAGCCTGAAAAACCTGTTCGTAGGGGGAAATCCGGGCTCCCGGTCGAGGGGTCGGACTTTTTGGACA>Bsp1286I|>BanII|>ApaI|>HaeIII>BglI|||>Sau96I|>Bsu36I>Bsp1286I>BsrSI|||||||>Sau96I|>DdeI>StyI|>HinfI||||||||||||3510|35203530|35403550CTGGGGCCCCTCCCTGAGGCTACAGTGCCCAAGGGGCAAGTTGGACTGGAGACCCCGGGGAGGGACTCCGATGTCACGGGTTCCCCGTTCAACCTGACCT>Bst71I>Sau96I||>Alw26I>AluI>BstOI||||||356035703580|3590|3600TTCCCAGCAGCCCCTCCCACTCCGAGACAAAATCAGCTACCCTGGGGCAGAAGGGTCGTCGGGGAGGGTGAGGCTCTGTTTTAGTCGATGGGACCCCGTC>HaeIII|>Sau96I||>BglI||>BstOI||||>HaeIII>SfiI|>BstOI|||||||3610||362036303640|GGCCTCATTGGCCCCAGGAAACCCCAGCCTGTCAGCACCTGTTCCAGGCCGGAGTAACCGGGGTCCTTTGGGGTCGGACAGTCGTGGACAAGGTCC

F. BlastResult(mouseUPIIwithgenome)>ref|NT_039473.1|Mm9_39513_30Musmusculuschromosome9genomiccontig,strainC57BL/6JLength=13248754Score=6837bits(3556),Expect=0.0Identities=3631/3663(99%),Gaps=21/3663(0%)Strand=Plus/MinusQuery:6ggatctcggccctctttctgcatccttgtcctaaatcattttcatatcttgctagacctc65||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917756ggatctcggccctctttctgcatccttgtcctaaatcattttcatatcttgctagacctc3917697Query:66agtttgagagaaacgaaccttctcattttcaagttg‐‐‐aaaaaaaaaagaggttcaaag122|||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917696agtttgagagaaacgaaccttctcattttcaagttgaaaaaaaaaaaaagaggttcaaag3917637Query:123tggctcactcaaagttacaagccaacactcaccactacgagtacaatggccaccattagt182||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917636tggctcactcaaagttacaagccaacactcaccactacgagtacaatggccaccattagt3917577Query:183gtctggcatgccccaggagacaggcatgcatattattctagatgactgggaggcagaggg242||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917576gtctggcatgccccaggagacaggcatgcatattattctagatgactgggaggcagaggg3917517Query:243gtggcctagtgaggtcagactgtggacagatcaggcagatgtgggttctgatcccaattc302||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917516gtggcctagtgaggtcagactgtggacagatcaggcagatgtgggttctgatcccaattc3917457Query:303ctcaggccgcagaactactgtggttcaagaaggggacaaaaggactgcagtcccggaaca362||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917456ctcaggccgcagaactactgtggttcaagaaggggacaaaaggactgcagtcccggaaca3917397Query:363ggaggtccactttgagagctgactgagcagaagaggaaagtgaagaacttcaggggcaag422|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917396ggaggtccgctttgagagctgactgagcagaagaggaaagtgaagaacttcaggggcaag3917337Query:423agcttaccctacttttacagctttgttgtcttctttactccaggggcgtccctggtactc482|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917336agcttaccctacttttacagctttgttgtcttctttactccaggggcatccctggtactc3917277Query:483agtaaatgtctgttggcttgaggaacatatgtgtaaggaggaaggagagggaacttgagg542||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917276agtaaatgtctgttggcttgaggaacatatgtgtaaggaggaaggagagggaacttgagg3917217Query:543gagttaagactcaagaatcaatcaaggagaggacagcagagaagacagggtttgggagag602||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917216gagttaagactcaagaatcaatcaaggagaggacagcagagaagacagggtttgggagag3917157Query:603agactccagacattggccctggttcccttcttggccactgtgaaaccctccagaggaact662||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct:3917156agactccagacattggccctggttcccttcttggccactgtgaaaccctccagaggaact3917097Query:663gagtgctgtggctttaaatgatctcagcactgtcagtgaagccgctctgctcaaagagtt722||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917096gagtgctgtggctttaaatgatctcagcactgtcagtgaagccgctctgctcaaagagtt3917037Query:723atcctcttgctcctgtgccgggg‐cctccccctcctctcagctccccaaacccttctcag781|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3917036atcctcttgctcctgtgccgggggcctccccctcctctcagctccccaaacccttctcag3916977Query:782ccactgtgatgggataattagatgcgagagctcagcacagatgatgctccagttgcttag841||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916976ccactgtgatgggataattagatgcgagagctcagcacagatgatgctccagttgcttag3916917Query:842caactaatggtttccatggagaccgcaaagcacagcctctagagcagccagtgagcagct901||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916916caactaatggtttccatggagaccgcaaagcacagcctctagagcagccagtgagcagct3916857Query:902cggcagggcagggagaagacgcaactctgctcctccagaaacctggggagggttagggtg961||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916856cggcagggcagggagaagacgcaactctgctcctccagaaacctggggagggttagggtg3916797Query:962gggaaggtgaggggggattggagggagatgggaaggcttaaaggcacaggaccctcatat1021||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916796gggaaggtgaggggggattggagggagatgggaaggcttaaaggcacaggaccctcatat3916737Query:1022ccctttaaaaatgtgaccagagatctgctctttttatcccacactgtctcacttctaatt1081||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916736ccctttaaaaatgtgaccagagatctgctctttttatcccacactgtctcacttctaatt3916677Query:1082tcaaatggggcttaggagtggacctatatggtctgctctgcagtaaaccaatcctcttcc1141||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916676tcaaatggggcttaggagtggacctatatggtctgctctgcagtaaaccaatcctcttcc3916617Query:1142tgattcctcctaaatcctcacgtaaacactgtgctatgtggaggtttgagacttgccagg1201||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916616tgattcctcctaaatcctcacgtaaacactgtgctatgtggaggtttgagacttgccagg3916557Query:1202aagatctcctgaccctgacaccctaccgtgtagccctcttccgcaccgccagttctccct1261||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916556aagatctcctgaccctgacaccctaccgtgtagccctcttccgcaccgccagttctccct3916497Query:1262cgtgtgtgatctttctgtgtgtaaagtgaggaggagatctcatctaaaggggacaaaagc1321||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916496cgtgtgtgatctttctgtgtgtaaagtgaggaggagatctcatctaaaggggacaaaagc3916437Query:1322caaggagctggaaaaacagaaatca‐ttc‐agttctccacccaaaggcagcctgagaaaa1379||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916436caaggagctggaaaaacagaaatcagttccagttctccacccaaaggcagcctgagaaaa3916377Query:1380acctctgtcccactttgcatttgcatgtgtgtgtgtgtatgagtgtgtgtgtgtatgagt1439

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916376acctctgtcccactttgcatttgcatgtgtgtgtgtgtatgagtgtgtgtgtgtatgagt3916317Query:1440gtgtgtgtgtatgagtgtgtgtgtgtatgagtgtgtgtgtgtatgagtgtgtgtgtgtgg1499||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916316gtgtgtgtgtatgagtgtgtgtgtgtatgagtgtgtgtgtgtatgagtgtgtgtgtgtgg3916257Query:1500atgagtgtgtgtgtgcacatgtatgagtgtgtctgtgtatgtatgattatatgtgtgtgt1559||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916256atgagtgtgtgtgtgcacatgtatgagtgtgtctgtgtatgtatgattatatgtgtgtgt3916197Query:1560ctgtgtgtgtgtatgagtatgtgtgtctgtgtgtgtgtatgagtatgtgtgtctgtgtgt1619||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916196ctgtgtgtgtgtatgagtatgtgtgtctgtgtgtgtgtatgagtatgtgtgtctgtgtgt3916137Query:1620gtggatgagtatgtgtgtctgtgtgtgtggatgagtgtgtgtgtggatgtgtatgaatat1679||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916136gtggatgagtatgtgtgtctgtgtgtgtggatgagtgtgtgtgtggatgtgtatgaatat3916077Query:1680gtgaatgtgagagtgcatgtgtgtgtgtctgtgtgtatacgagtatgtgtgtgcatgtgg1739||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916076gtgaatgtgagagtgcatgtgtgtgtgtctgtgtgtatacgagtatgtgtgtgcatgtgg3916017Query:1740ataagtgtgtgtgtgtgtgtatgagtatgtgtgtgtgtctgtgtatgagtgtttacacca1799||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3916016ataagtgtgtgtgtgtgtgtatgagtatgtgtgtgtgtctgtgtatgagtgtttacacca3915957Query:1800atacagcatctgctaactccaaatgtgagtctgtgtctctgcgttacctgtaagcgtcat1859||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915956atacagcatctgctaactccaaatgtgagtctgtgtctctgcgttacctgtaagcgtcat3915897Query:1860aggaagttttacgaaaccatgagatggctcaacaggtaaaagctcttgccaccatgcctg1919||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915896aggaagttttacgaaaccatgagatggctcaacaggtaaaagctcttgccaccatgcctg3915837Query:1920acattttggtccccagaagccacattatgaaaggagagagccaatgcccacaacttgccc1979||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915836acattttggtccccagaagccacattatgaaaggagagagccaatgcccacaacttgccc3915777Query:1980tctgacctccacatagacactgtggcacacatatgggcacacatataaaataataaatgc2039||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915776tctgacctccacatagacactgtggcacacatatgggcacacatataaaataataaatgc3915717Query:2040aactttaaatttttgaaagaaattaagtttcgggttttacctcagacccactgtatcaga2099||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915716aactttaaatttttgaaagaaattaagtttcgggttttacctcagacccactgtatcaga3915657Query:2100agctacatctgaaccagtttcctgatcgtccattgcacagtaaatttggagatcgtctgg2159||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915656agctacatctgaaccagtttcctgatcgtccattgcacagtaaatttggagatcgtctgg3915597

Query:2160tgagagttcctcctgctcccagccggagcccccaggttctgttgtcttctaccccctaca2219||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915596tgagagttcctcctgctcccagccggagcccccaggttctgttgtcttctaccccctaca3915537Query:2220cacatcgcagactttaagtccacccagcatgggggaaaggacagaaccccatcagccctt2279|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915536cacattgcagactttaagtccacccagcatgggggaaaggacagaaccccatcagccctt3915477Query:2280cagtggcccccacttttccactaggacctttctctggggttggggaaaatgggtggttcc2339||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915476cagtggcccccacttttccactaggacctttctctggggttggggaaaatgggtggttcc3915417Query:2340tggagcagaccctgacggtctgagctcaggtcctatcgagttcacctagctgagacaccc2399||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915416tggagcagaccctgacggtctgagctcaggtcctatcgagttcacctagctgagacaccc3915357Query:2400acacccctgcagccactttgcagtgacaacctgagtctcaggttctgcatctataaaaag2459||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915356acacccctgcagccactttgcagtgacaacctgagtctcaggttctgcatctataaaaag3915297Query:2460cagtgcctttcaggagggcatgcagagccccctggccagcgtctagaggagaggtgactg2519||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915296cagtgcctttcaggagggcatgcagagccccctggccagcgtctagaggagaggtgactg3915237Query:2520agtggggccatgtcactcgtccatggctggagaacctccatcagtttctcccagttagcc2579||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915236agtggggccacgtcactcgtccatggctggagaacctccatcagtt‐ctcccagttagcc3915178Query:2580tggggcaggagagaaccagaggagctgtggctgctgatggtgtttcttccccaactgttg2639||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915177tggggcaggagagaaccagaggagctgtggctgctgatggtgtttcttccccaactgttg3915118Query:2640tcccaggcatcgaaccccagagcgacctgcacacagtccaccgctgagccacaccccagc2699||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915117tcccaggcatcgaaccccagagcgacctgcacacagtccaccgctgagccacaccccagc3915058Query:2700cctccacctcctctgctcctggttacaggattgtttt‐gtcttgaagggttttgttgttg2758|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3915057cctccacctcctctgctcctggttacaggattgtttttgtcttgaagggttttgttgttg3914998Query:2759ctactttttgctttgttttt‐t‐‐‐‐‐cttttttctttttt‐‐‐‐‐‐‐aacataaggttt2805||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914997ctactttttgctttgtttttgtttttgcttttttttttttttttttttaacataaggttt3914938Query:2806ctctgtgtagccctagctgtcctggaactcactctgtagaccaggctggcctcaaactca2865|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914937ctctttgtagccctagctgtcctggaactcactctgtagaccaggctggcctcaaactca3914878Query:2866gaaatccaccttcctcccaagtgctgggattaaaggcattcgccaccatcgcccagcccc2925||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914877gaaatccaccttcctcccaagtgctgggattaaaggcattcgccaccatcgcccagcccc3914818

Query:2926cggtcttgtttcctaaggttttcctgctttactcgctacccgttgcacaaccgcttgctg2985|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914817tggtcttgtttcctaaggttttcctgctttactcgctacccgttgcacaaccgcttgctg3914758Query:2986tccaagtctgtttgtatctactccaccgcccactagccttgctggactggacctacgttt3045||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914757tccaagtctgtttgtatctactccactgcccactagccttgctggactggacctgcgttt3914698Query:3046acctggaagccttcactaacttcccttgtctccaccttctggagaaatctgaaggctcac3105||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914697acctggaagccttcactaacttcccttgtctccaccttctggagaaatctgaaggctcac3914638Query:3106actgataccctccgcttctcccagagtcgcagtttcttaggcctcagttaaataccagaa3165|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914637actgataccctccgcttctcccagagtcgcagtttcttaggcctcagttaaatactagaa3914578Query:3166ttggatctcaggctctgctatccccaccctacctaaccaaccccctcctctcccatcctt3225||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914577ttggatctcaggctctgctatccccaccctacctaaccaaccccctcctctcccatcctt3914518Query:3226actagccaaagccctttcaacccttggggcttttcctacacctacacaccagggcaattt3285||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914517actagccaaagccctttcaacccttggggcttttcctacacctacacaccagggcaattt3914458Query:3286tagaactcatggctctcctaggaaaacgcctacctccttggagacagcaccctctacagt3345||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914457tagaactcatggctctcctaggaaaacgcctacctccttggagacagcaccctctacagt3914398Query:3346ccaggaggcagacactcagacagaggaactctgtccttcagtcgcgggagttccagaaag3405||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914397ccaggaggcagacactcagacagaggaactctgtccttcagtcgcgggagttccagaaag3914338Query:3406agccatactcccctgcagctaactaagctgccaggacccagccagagcatccccctttag3465||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914337agccatactcccctgcagctaactaagctgccaggacccagccagagcatccccctttag3914278Query:3466gcccgagggccagctccccagcctgaaaaacctgtctggggcccctccctgaggctacag3525||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914277gcccgagggccagctccccagcctgaaaaacctgtctggggcccctccctgaggctacag3914218Query:3526tgcccaaggggcaagttggactggattcccagcagcccctcccactccgagacaaaatca3585||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914217tgcccaaggggcaagttggactggattcccagcagcccctcccactccgagacaaaatca3914158Query:3586gctaccctggggcagggcctcattggccccaggaaaccccagcctgtcagcacctgttcc3645|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||Sbjct:3914157gctatcctggggcagggcctcattggccccaggaaaccccagcctgtcagcacctgttcc3914098Query:3646agg3648|||Sbjct:3914097agg3914095

G. Vectormaphttp://www.stratagene.com/vectors/selection/plasmid1.htm