《PET质粒图谱大全》

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1、pET-3a-dVectorsTB02612/98ThepET-3a-dvectorscarryanN-terminalT7•Tag®sequenceandBamHIcloningsite.TheseCat.No.pET-3aDNA69418-3vectorsaretheprecursorstomanypETfamilyvectors;thepET-23a-d(+)seriescorrespondstopET-3bDNA69419-3pET-3a-dbutincorporatesseveraladditionalfeatures.Un

2、iquesitesareshownonthecirclemap.pET-3cDNA69420-3NotethatthesequenceisnumberedbythepBR322convention,sotheT7expressionregionispET-3dDNA69421-3reversedonthecircularmap.Thecloning/expressionregionofthecodingstrandtranscribedbyT7RNApolymeraseisshownbelow.pET-3asequencelandma

3、rksEcoRI(4638)T7promoter615-631ApoI(4638)T7transcriptionstart614ClaI(24)HindIII(29)T7•Tagcodingsequence519-551AatII(4567)Bpu1102I(458)T7terminator404-450SspI(4449)BamHI(510)pBR322origin2814ScaI(4125)blacodingsequence3575-4432NdeI(550)PvuI(4015)XbaI(588)2)ThemapsforpET-3

4、b,pET-3candpET-3d43BglII(646)PstI(3890)5-4SgrAI(687)arethesameaspET-3a(shown)withthe75followingexceptions:pET-3bisa4639bp(3SphI(843)plasmid;subtract1bpfromeachsitebeyondpAEcoNI(903)BamHIat510.pET-3cisa4638bpplasmid;SalI(928)subtract2bpfromeachsitebeyondBamHIEam1105I(364

5、5)–PshAI(993)at510.pET-3disa4637bpplasmid;theBamHIsiteisinthesamereadingframeasinpET-3c.AnNcoIsiteissubstitutedforpET-3aEagI(1216)(4640bp)theNdeIsitewithanet1bpdeletionatNruI(1251)position550ofpET-3c.Asaresult,NcoIHgiEII(3338)ApaBI(1329)cutspET-3dat546.Fortherestofthesi

6、tes,BspMI(1331)subtract3bpfromeachsitebeyondpositionAlwNI(3168)551inpET-3a.NdeIdoesnotcutpET-3d.ori(281BsmI(1636)4)AvaI(1702)MscI(1723)BspLU11I(2752)Bpu10I(1858)AflIII(2752)BscGI(1912)SapI(2636)Bst1107I(2523)BsaAI(2504)Tth111I(2497)BsmBI(2393)PvuII(2343)T7promoterprimer

7、#69348-3T7promoterBglIIXbaIrbsAGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGAGACCACAACGGTTTCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGANdeIT7•TagpET-3aBamHIBpu1102ITATACATATGGCTAGCATGACTGGTGGACAGCAAATGGGTCGCGGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAAMe

8、tAlaSerMetThrGlyGlyGlnGlnMetGlyArgGlySerGlyCysEndT7terminatorprimer#69337-3pET-3b...GGTCGGGATCCGGCTGCTAACAAAGC