pET-30a DNApET-30b DNApET-30c DNA
Cat. No.69909-369910-369911-3
TB095 12/98
The pET-30a-c(+) vectors carry an N-terminal His•Tag®/thrombin/S•Tag™/enterokinase config-uration plus an optional C-terminal His•Tag sequence. Unique sites are shown on the circle map.Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is re-versed on the circular map. The cloning/expression region of the coding strand transcribed by T7RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage willproduce virions containing single-stranded DNA that corresponds to the coding strand. Therefore,single-stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).
pET-30a(+) sequence landmarks
Pvu I(4479)Sgf I(4479)Sma I(4353)The maps for pET-30b(+) and pET-30c(+)are the same as pET-30a(+) (shown) withthe following exceptions: pET-30b(+) is a5421bp plasmid; subtract 1bp from each sitebeyond BamH I at 198. pET-30c(+) is a5423bp plasmid; add 1bp to each sitebeyond BamH I at 198.
kan (4048-4860)T7 promoter
T7 transcription startHis•Tag coding sequenceS•Tag coding sequenceMultiple cloning sites(NcoI - XhoI)
His•Tag coding sequenceT7 terminator
lacIcoding sequencepBR322 origin
Kan coding sequencef1 origin
419-435418327-344249-293158-217140-15726-72826-19053339
4048-48604956-5411
Dra III(5180)Xho I(158)Not I(166)Eag I(166)Hind III(173)Sal I(179)Sac I(190)EcoR I(192)BamH I(198)EcoR V(206)Nco I(212)Kpn I(238)Bgl II(241)Bpu1102 I(80)Nsp V(268)Nde I(346)Xba I(384)SgrA I(495)Sph I(651)56-5411) (49nigori 1fMlu I(1176)Bcl I(1190)BstE II(1357)Apa I(1387)lacI (826-1905)Nru I(4136)pET-30a(+)(5422bp)Eco57 I(3825)AlwN I(3693)BssH II(1587)Hpa I(1682)orBssS I(3450)(3BspLU11 I(3277)Sap I(3161)Bst1107 I(3048)Tth111 I(3022)i 339)PshA I(2021)Bgl I(2240)Fsp I(2258)Psp5 II(2283)T7 promoter primer #69348-3pET upstream primer #69214-3lac operatorrbsXba IAGATCGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGAS•TagNsp VHis•TagBgl IINde ITATACATATGCACCATCATCATCATCATTCTTCTGGTCTGGTGCCACGCGGTTCTGGTATGAAAGAAACCGCTGCTGCTAAATTCGAACGCCAGCACATGGACAGCCCAGATCTG MetHisHisHisHisHisHisSerSerGlyLeuValProArgGlySerGlyMetLysGluThrAlaAlaAlaLysPheGluArgGlnHisMetAspSerProAspLeuthrombinEag IpET-30a(+)His•TagKpn INco IEcoR VBamH IEcoR ISac INot IXho ISal IHind IIIGGTACCGACGACGACGACAAGGCCATGGCTGATATCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAAGlyThrAspAspAspAspLysAlaMetAlaAspIleGlySerGluPheGluLeuArgArgGlnAlaCysGlyArgThrArgAlaProProProProProLeuArgSerGlyCysEndenterokinasepET-30b(+) ...GCGATATCGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGA... AlaIleSerAspProAsnSerSerSerValAspLysLeuAlaAlaAlaLeuGluHisHisHisHisHisHisEndpET-30c(+) ...GGATATCTGTGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAA... ...GlyTyrLeuTrpIleArgIleArgAlaProSerThrSerLeuArgProHisSerSerThrThrThrThrThrThrGluIleArgLeuLeu...T7 terminatorBpu1102 ICAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGT7 terminator primer #69337-3T7 promoterpET-30a-c(+) cloning/expression regionNovagen •ORDERING800-526-7319 •
TECHNICALSUPPORT
800-207-0144
pET-30a(+) Restriction Sites
EnzymeAccIAceIII# SitesLocations218030477943167132295020AciI75AflIII211763277AluI22AlwI13Alw21I715919030953595Alw44I311563091AlwNI13693ApaI11387ApaBI1860ApoI719227049824993AvaI21584351AvaII517282104BamHI1198BanI1023431010961815BanII61905605255BbsI413221661BbvI25BccI14Bce83I62119903907BcefI669510365206BcgI816019420362854BclI11190BfaI6703855331BglI12240BglII1241BmgI11385BpmI410141503Bpu10I223834496Bpu1102I180BsaAI230295180BsaBI3449459BsaHI5499520BsaJI105721222493437BsaWI72149536304614BsaXI218355128BsbI229935087BscGI11BsgI310271227BsiI13450BsiEI51691961BslI26BsmI243634440BsmAI687312784495BsmBI317912918BsmFI46372178BsoFI43Bsp24I1046649813513770Bsp1286I12BspEI222466BspGI12803BspLU11I13277BsrI21BsrBI44053210BsrDI212231589BsrFI748649544335281BssHII11587200227862927EnzymeBst1107IBstEIIBstXIBstYICac8ICjeICjePIClaICviJICviRIDdeIDpnIDraIIIDrdIDrdIIDsaIEaeIEagIEarIEciIEco47IIIEco57IEcoNIEcoO109IEcoRIEcoRIIEcoRVFauIFokIFspIGdiIIHaeIHaeIIHaeIIIHgaIHgiEIIHhaIHin4IHincIIHindIIIHinfIHpaIHphIKpnIMaeIIMaeIIIMboIIMluIMmeIMnlIMseIMslIMspIMspA1I13193980MwoINarINciINcoINdeINgoAIVNlaIIINlaIVNotINruINsiI# Sites11394024182853111231323413331231911791471424112464211811611416131725256299394121142623112Locations3048135797811071321982469391812302413929EnzymeNspINspVPfl1108IPflMIPleIPshAIPsp5IIPsp1406IPvuIPvuIIRcaIRsaISacISalISapISau96ISau3AIScrFISfaNISfcISgfISgrAISmaISphISspIStyITaqITaqIITfiIThaITseITsp45ITsp509ITth111ITth111II774203181173168223838631075168241654707UbaJIVspIXbaIXcmIXhoIXmnI1851312# Sites4113911413341111423212341111221769362572118Locations651262226820632607584337253656471120212283838220644791776186957439972361323190179316129143281TB095 12/98
74047281952474281251151608512331716763591116022714534170260228684872308349654314145140924276219249819455742035228351920711387239525626335217413421601663146828882291336837791502366647982002518029708992121661667949535813825711531928993424206122228222258166217375533855185613484316133512082439160912143437513522496164292349725311850228317544367181147243303418447949543536514404571084508418554390354237335399497221213022157444619752327461831792831470947333252123129634849044566249641176162225255847231850329226363772407933031357462630221015390643338410321582835218553532716292930242137280417084315186115484868273844421920156638673874247463461343501998467848671133619435124661816181147523483243731933617447911763492468636765157412143154677Enzymes that do not cut pET-30a(+):AatIIAflIIAgeIAscIBaeIBsaIBseRIBspMIBsu36IDraIEam1105IFseIMunINheIPacIPmeIPstIRleAIRsrIISacIISexAISfiISnaBISpeISse8387IStuISunISwaIAvrIIBsrGIMscIPmlIScaISrfI140444952548101738021791291812282850842868499212346486151615462264245953951049394828329875201206361963417763864181618692074223452814878862532420742234166413643294595Novagen •FAX608-238-1388 •E-MAILnovatech@novagen.com
因篇幅问题不能全部显示，请点此查看更多更全内容