PlasmidInfo:
PlasmidInformation
ProductName:pSF-CMV-PGK
ProductCode:OG387
Size(bp):4747bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:Cytomegalovirus(CMV)immediateearlypromoter/MousePhosphoglycerateKinase(PGK)Promoter
PlasmidPurpose:
Thisplasmidcontainstwopromotersthatterminatetranscriptionatthesamepoly-adenylationsignalallowingtheexpressionoftwogenesfromoneexpressioncassette.Thesecondpromoter(PGK)isapproximately10-foldweakerthantheupstreamCMVpromoter.
PromoterExpressionLevel:ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.HowevertherearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.ForthisreasonwestockarangeofotherpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.
SequenceandMap:
OtherInfo:
TranscriptionTermination:Thisplasmidcontainsthreealternativetranscriptionterminatorsformammalianbacterialandbacteriophage(T7)expression.Thismeansthatonlythepromoterneedstobechangedtoaltertheexpressionsystemyouareusing.Wesellmultiplepromotersthatcanbeusedineachofthesesystems.Thepresenceofeachterminatordoesnotreduceexpressioninthealternativesystems.
Cloning:
MultipleCloningSiteNotes:ThereisastartcodonintheNcoIsitethatcanberemovedbydigestionwithKpnIifrequired.TheMCSforgeneinsertionsextendsfromNotItoXbaIbuttheShine-DalgarnosequencesandKOZAKribosomalentrysitesequencesarealignedwiththestartcodonintheNcoIsite.ThePGKpromoterhasbeeninsertedbetweentheClaIandBamHIsitesandcontainssitesdownstreamtoallowasecondgenetobeinserted.
TheBsgIandBseRIrestrictionsitesinthefirstMCScleavewithinthestopcodonintheXbaIsiteandallowtheretrospectivefusionofcodingsequences.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
