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Oxford Genetics/pSF-CMV-COOH-EKT-V5-2 (OG61) C-Terminal V5 Tag Plasmid/OG61/1 Ea
PlasmidInfo:
PlasmidInformation
ProductName:pSF-CMV-COOH-EKT-V5-2
ProductCode:OG61
Size(bp):4284bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:Cytomegalovirus(CMV)immediateearlypromoter
PlasmidPurpose:
ThisvectoraddsaV5tagthatallowsproteindetectionorpurificationusingantibodiesagainsttheV5peptide.TheV5tagwillbepositionedatthe3primeend(Cterminus)ofagene(expressedprotein)thatiswithintheprimaryMCS(NotItoXbaI).TheV5tagcodingsequenceisGKPIPNPLLGLDST.Thereisanenterokinasecleavagesiteimmediatelyupstreamofthistag(DDDDK)thatcanbeusedtoremovethetagfromapurifiedproteinifrequired.Itcleavesafterthelysineresidue.
PromoterExpressionLevel:ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.HowevertherearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.ForthisreasonwestockarangeofotherpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.
SequenceandMap:
OtherInfo:
TranscriptionTermination:Thisplasmidcontainsthreealternativetranscriptionterminatorsformammalianbacterialandbacteriophage(T7)expression.Thismeansthatonlythepromoterneedstobechangedtoaltertheexpressionsystemyouareusing.Wesellmultiplepromotersthatcanbeusedineachofthesesystems.Thepresenceofeachterminatordoesnotreduceexpressioninthealternativesystems.
Cloning:
CloninginaGene:ThereisastartcodonintheNcoIsitecanberemovedbydigestionwithKpnIifrequired.TheMCSforgeneinsertionsextendsfromNotItoXbaI.TherearebuiltinShine-DalgarnoandKOZAKsequencesalignedwiththestartcodonwithintheNcoIrestrictionsite.Iftheseareremovedyouwillneedtoaddaribosomalentrysitetothe5primeendofyourgeneforoptimalexpression.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
