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Oxford Genetics/pSF-CMV-COOH-FLAG2 (OG54) C-Terminal FLAG® Tag Plasmid/OG54/1 Ea
PlasmidInfo:
PlasmidInformation
ProductName:pSF-CMV-COOH-FLAG2
ProductCode:OG54
Size(bp):4251bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:Cytomegalovirus(CMV)immediateearlypromoter
PlasmidPurpose:
ThisvectoraddsaFLAGtagthatallowsproteindetectionorpurificationusingantibodiesagainstthepeptide.TheFLAGtagwillbepositionedatthe3primeend(Cterminus)ofagene(expressedprotein)intheprimaryMCS(NotItoXbaI).ForreferencethecodingsequenceoftheFLAGtagisonebasepairoutofframewiththeTAGstopcodonthatisfoundwithintheupstreamXbaIsiteinpSF-CMV.TheFLAGtagcodingsequenceisDYKDDDDK.Thissequencecontainsanenterokinasecleavagesequence(DDDDK)howeverwhenusedasaC-terminaltagthiswillnotremovethetagbecauseitcleavesafterthefinallysinewhichwillbethefinalaminoacidinanyproteinproducedfromthisvector.
PromoterExpressionLevel:ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.HowevertherearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.ForthisreasonwestockarangeofotherpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.
FLAGisaregisteredtrademarkoftheSigmaAldrichCorporation.
SequenceandMap:
OtherInfo:
TranscriptionTermination:Thisplasmidcontainsthreealternativetranscriptionterminatorsformammalianbacterialandbacteriophage(T7)expression.Thismeansthatonlythepromoterneedstobechangedtoaltertheexpressionsystemyouareusing.Wesellmultiplepromotersthatcanbeusedineachofthesesystems.Thepresenceofeachterminatordoesnotreduceexpressioninthealternativesystems.
Cloning:
CloninginaGene:ThereisastartcodonintheNcoIsitecanberemovedbydigestionwithKpnIifrequired.TheMCSforgeneinsertionsextendsfromNotItoXbaI.TherearebuiltinShine-DalgarnoandKOZAKsequencesalignedwiththestartcodonwithintheNcoIrestrictionsite.Iftheseareremovedyouwillneedtoaddaribosomalentrysitetothe5primeendofyourgeneforoptimalexpression.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
