pET28质粒图谱

Novagen•ORDERING800-526-7319•TECHNICALSUPPORT800-207-0144AGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACCATGGGCAGCAGCCATCATCATCATCATCACAGCAGCGGCCTGGTGCCGCGCGGCAGCCATATGGCTAGCATGACTGGTGGACAGCAAMetGlySerSerHisHisHisHisHisHisSerSerGlyLeuValProArgGlySerHisMetAlaSerMetThrGlyGlyGlnGlnATGGGTCGCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCCpET-28a(+)MetGlyArgGlySerGluPheGluLeuArgArgGlnAlaCysGlyArgThrArgAlaProProProProProLeuArgSerGlyCysEnd...GGTCGGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCCpET-28b(+)...GlyArgAspProAsnSerSerSerValAspLysLeuAlaAlaAlaLeuGluHisHisHisHisHisHisEnd...GGTCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCCpET-28c(+)...GlyArgIleArgIleArgAlaProSerThrSerLeuArgProHisSerSerThrThrThrThrThrThrGluIleArgLeuLeuThrLysPro...GAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGBglIINheIXbaIlacoperatorpETupstreamprimer#69214-3T7promoterprimer#69348-3T7promoterrbsBpu1102IT7terminatorprimer#69337-3T7terminatorT7•TagHis•TagthrombinNdeIHis•TagHindIIIEcoRIXhoIBamHISacINotINcoIEagISalIpET-28a-c(+)cloning/expressionregionTB07412/98ThepET-28a-c(+)vectorscarryanN-terminalHis•Tag®/thrombin/T7•Tag®configurationplusanoptionalC-terminalHis•Tagsequence.Uniquesitesareshownonthecirclemap.NotethatthesequenceisnumberedbythepBR322convention,sotheT7expressionregionisreversedonthecircularmap.Thecloning/expressionregionofthecodingstrandtranscribedbyT7RNApoly-meraseisshownbelow.Thef1originisorientedsothatinfectionwithhelperphagewillproducevirionscontainingsingle-strandedDNAthatcorrespondstothecodingstrand.Therefore,single-strandedsequencingshouldbeperformedusingtheT7terminatorprimer(Cat.No.69337-3).pET-28a(+)sequencelandmarksT7promoter370-386T7transcriptionstart369His•Tagcodingsequence270-287T7•Tagcodingsequence207-239Multiplecloningsites(BamHI-XhoI)158-203His•Tagcodingsequence140-157T7terminator26-72lacIcodingsequence773-1852pBR322origin3286Kancodingsequence3995-4807f1origin4903-5358ThemapsforpET-28b(+)andpET-28c(+)arethesameaspET-28a(+)(shown)withthefollowingexceptions:pET-28b(+)isa5368bpplasmid;subtract1bpfromeachsitebeyondBamHIat198.pET-28c(+)isa5367bpplasmid;subtract2bpfromeachsitebeyondBamHIat198.pET-28a-c(+)VectorslacI(773-1852)ori(3286)Kan(3995-4807)f1origin(4903-5358)Bpu1102I(80)XbaI(335)BglII(401)SgrAI(442)SphI(598)MluI(1123)BclI(1137)BstEII(1304)ApaI(1334)BssHII(1534)EcoRV(1573)HpaI(1629)PshAI(1968)BglI(2187)FspI(2205)Psp5II(2230)Tth111I(2969)Bst1107I(2995)SapI(3108)BspLU11I(3224)BssSI(3397)AlwNI(3640)Eco57I(3772)NruI(4083)ClaI(4117)SmaI(4300)PvuI(4426)SgfI(4426)DraIII(5127)XhoI(158)NotI(166)EagI(166)HindIII(173)SalI(179)SacI(190)EcoRI(192)BamHI(198)NheI(231)NdeI(238)NcoI(296)pET-28a(+)(5369bp)Cat.No.pET-28aDNA69864-3pET-28bDNA69865-3pET-28cDNA69866-3Call137-649-32037tobuythevectorinChina.TB07412/98Enzyme#SitesLocationsAccI21802994AceIII7890161819492733287431764967AciI77AflIII211233224AluI22AlwI13Alw21I71591906231107221830423542Alw44I3110330383538AlwNI13640ApaI11334ApaBI1807ApoI619213984039422349294940AvaI21584298AvaII516752051213922302509BamHI1198BanI925344546658010431762189220185164BanII6190507521133440815202BbsI41269160819822342BbvI27BccI14Bce83I62119372107331536133854BcefI66429831610372647455153BcgI9160194228141514491949198328012835BclI11137BfaI7702323362238371940265278BglI12187BglII1401BmgI11332BpmI4961145020842751Bpu10I223304443Bpu1102I180BsaAI229765127BsaBI34004062421BsaHI544646758110801763BsaJI1057296560566175821963384429742984699BsaWI72144219452413343035774561BsaXI217825075BsbI229405034BscGI11BsgI397411742384BsiI13397BsiEI51691908314035644426BslI23BsmI243104387BsmAI682012251351173828654442BsmBI3173828654442BsmFI4584212524955342BsoFI48Bsp24I12Bsp1286I12BspEI222413BspGI12750BspLU11I13224BsrI22BsrBI4356315748255271BsrDI211701536BsrFI74334428092021218143805228BssHII11534Bst1107I12995Enzyme#SitesLocationsBstEII11304BstXI392510541177BstYI913219840168718992416386538764675Cac8I40CjeI26CjePI30ClaI14117CviJI86CviRI22DdeI11DpnI21DraIII15127DrdI3291733325082DrdII28465132DsaI32965602196EaeI41664315631797EagI1166EarI374131084239EciI390032983444Eco47III352820292478Eco57I13772EcoNI26584338EcoO109I3535562230EcoRI1192EcoRII1025684611611701175832503371338443144671EcoRV11573FauI17FokI9116911782443250525832769291040644670FspI12205GdiII41664315631797HaeI685121723239325037024513HaeII14HaeIII24HgaI11HgiEII27213810HhaI47Hin4I3102241124654HincII21811629HindIII1173HinfI18HpaI11629HphI16MaeII14MaeIII16MboII12MluI11123MmeI73439362340684262462446335104MnlI25MseI25MslI6117514631493221124062797MspI29MspA1I9842641153172318162815293435663811MwoI39NarI44464675811763NciI12NcoI1296NdeI1238NgoAIV4433202121815228NheI1231NlaIII26NlaIV22NotI1166NruI14083NsiI242764542NspI4598256928613228Enzyme#SitesLocationsPfl1108I12010PflMI27054689PleI9384672759155531183603465850625070PshAI11968Psp5II12230Psp1406I4785215325494912PvuI14426PvuII31723181