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Novagen•ORDERING800-526-7319•TECHNICALSUPPORT800-207-0144AGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGAGACCACAACGGTTTCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGGCTAGCATGACTGGTGGACAGCAAATGGGTCGCGGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAAMetAlaSerMetThrGlyGlyGlnGlnMetGlyArgGlySerGlyCysEndpET-3b...GGTCGGGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAA...GlyArgAspProAlaAlaAsnLysAlaArgLysGluAlaGluLeuAlaAlaAlaThrAlaGluGlnEnd...TACCATGGCTAGC...pET-3c,d...GGTCGGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAAMetAlaSer......GlyArgIleArgLeuLeuThrLysProGluArgLysLeuSerTrpLeuLeuProProLeuSerAsnAsnEndCCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGNcoIpET-3dNdeIT7•TagT7terminatorBamHIBpu1102IT7terminatorprimer#69337-3pET-3arbsBglIIXbaIT7promoterprimer#69348-3T7promoterThepET-3a-dvectorscarryanN-terminalT7•Tag®sequenceandBamHIcloningsite.ThesevectorsaretheprecursorstomanypETfamilyvectors;thepET-23a-d(+)seriescorrespondstopET-3a-dbutincorporatesseveraladditionalfeatures.Uniquesitesareshownonthecirclemap.NotethatthesequenceisnumberedbythepBR322convention,sotheT7expressionregionisreversedonthecircularmap.Thecloning/expressionregionofthecodingstrandtranscribedbyT7RNApolymeraseisshownbelow.pET-3asequencelandmarksT7promoter615-631T7transcriptionstart614T7•Tagcodingsequence519-551T7terminator404-450pBR322origin2814blacodingsequence3575-4432ThemapsforpET-3b,pET-3candpET-3darethesameaspET-3a(shown)withthefollowingexceptions:pET-3bisa4639bpplasmid;subtract1bpfromeachsitebeyondBamHIat510.pET-3cisa4638bpplasmid;subtract2bpfromeachsitebeyondBamHIat510.pET-3disa4637bpplasmid;theBamHIsiteisinthesamereadingframeasinpET-3c.AnNcoIsiteissubstitutedfortheNdeIsitewithanet1bpdeletionatposition550ofpET-3c.Asaresult,NcoIcutspET-3dat546.Fortherestofthesites,subtract3bpfromeachsitebeyondposition551inpET-3a.NdeIdoesnotcutpET-3d.pET-3a-dcloning/expressionregionpET-3a-dVectorsTB02612/98pET-3a(4640bp)ori(2814)Ap(3575-4432)ClaI(24)HindIII(29)Bpu1102I(458)BamHI(510)NdeI(550)XbaI(588)BglII(646)SgrAI(687)SphI(843)EcoNI(903)SalI(928)PshAI(993)EagI(1216)NruI(1251)ApaBI(1329)BspMI(1331)BsmI(1636)AvaI(1702)MscI(1723)Bpu10I(1858)BscGI(1912)PvuII(2343)BsmBI(2393)Tth111I(2497)BsaAI(2504)Bst1107I(2523)SapI(2636)AflIII(2752)BspLU11I(2752)AlwNI(3168)HgiEII(3338)PstI(3890)PvuI(4015)ScaI(4125)SspI(4449)AatII(4567)EcoRI(4638)ApoI(4638)Eam1105I(3645)–Cat.No.pET-3aDNA69418-3pET-3bDNA69419-3pET-3cDNA69420-3pET-3dDNA69421-3Enzyme#SitesLocationsAatII14567AccI29292522AceIII59742261240227043944AciI86AflIII12752AluI18AlwI14Alw21I8280868145517462570307042314316Alw44I3256630664312AlwNI13168ApaBI11329ApoI14638AvaI11702AvaII810761164141317161758203737834005BamHI1510BanI9761196907118251043148215663593BanII2752766BbsI3100718704623BbvI24BccI97378301267135616631675368238064093Bce83I739996211322843314133824250BcefI388714443254BcgI850654097410082329236341504184BfaI82304485445891766324735003835BglI3121214463765BglII1646BpmI41109166322793715Bpu10I11858Bpu1102I1458BsaI26133706BsaAI12504BsaBI36456511949BsaHI6691712826148341824564BsaJI91151294358058111444164617242912BsaWI63809701941295831053936BsbI224684188BscGI11912BsiI3292543094616BsiEI728993312192668309240154164BslI21BsmI11636BsmAI4613239337064482BsmBI12393BsmFI4829115013752023BsoFI45Bsp24I10513545658690324532773423345545494581Bsp1286I10280752766868145517462570307042314316BspEI23801941BspGI3133614132278BspLU11I12752BspMI11331BsrI20BsrBI226854486BsrDI237063880BsrFI71606786871046120615603725Bst1107I12523BstYI95106461944339334043490350242704287Enzyme#SitesLocationsCac8I32CjeI18CjePI22ClaI124CviJI79CviRI21DdeI1045847918582020256030273436360241424568DpnI25DraI224452860DsaI28051724EaeI6295676808121617214033EagI11216Eam1105I13645EarI226364440EciI41672282629723800Eco47III423477310542006Eco57I233004312EcoNI1903EcoO109I5431801171617584621EcoRI14638EcoRII612913351718277828992912EcoRV2187378FauI11FokI12FspI4262163517333867GdiII529567680812164033HaeI71197126913261723276727783230HaeII11HaeIII23HgaI11HgiEII13338HhaI31Hin4I516334141836443718HincII29304186HindIII129HinfI11HphI12MaeII101178123418231847207725033455387142444564MaeIII17MboII11MmeI422230929673151MnlI30MscI11723MseI18MslI71308173919342325389740564415MspI28MspA1I7462141823432462309433394280MwoI37NarI46917128261483NciI1017181215361762209023962431313238284179NdeI1550NgoAIV4678104612061560NheI2229543NlaIII27NlaIV25NruI11251NspI4843209723892756Pfl1108I210353663PflMI215981647PleI5629917264631313634PshAI1993Psp5II217161758Psp1406I41178207738714244PstI13890Enzyme#SitesLocationsPvuI14015PvuII12343RcaI4766347244804585RsaI316525584125SalI1928SapI12636Sau96I16Sau3AI25ScaI14125ScrFI16SfaNI22SfcI5138614301732083886SgrAI1687SphI1843SspI14449StyI24351646TaqI9243396436519291404154528524296TaqII694726543993417843314348TfiI6112912831581180223062727ThaI25TseI24Tsp45I91242121157142421912404249939014112Tsp509I1058251580630159616103512381840734638Tth111I12497Tth111II522133342334933814637UbaJI21VspI26293817XbaI1588XmnI223104244EnzymesthatdonotcutpET-3a:Afl
本文标题:PET质粒图谱大全
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