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Oxford Genetics/pSF CAG BetaGal Ub Puro（OG506）CAG报告基因（β-Gal）质粒/OG506/1 Ea
PlasmidInfo:
PlasmidInformation
ProductName:pSF-CAG-BetaGal-Ub-Puro
ProductCode:OG506
Size(bp):10450bp
BacterialAntibioticSelection:AmpR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:CAGsyntheticmammalianpromoterandUbiquitinpromoter/ChickenBetaActin(CBA)promoter/HumanUbiquitinPromoter
PlasmidPurpose:
Thisplasmidisdesignedforcreatingbetagalactosidasestablecelllinesorthetransientexpressiononbetagalactosidase.ItcontainsthestrongconstitutivemammalianCAGpromotertodrivetheexpressionofthebetagalactosidasereporterandapuromycinresistancegenetoallowforstablecelllineselection.
PromoterExpressionLevel:ThisplasmidcontainsthemammalianCAGpromoterwhichisasyntheticcompositeoftheCMVimmediateearlyenhancerfollowedbytheCBApromoterandtherabbitbetaglobinintron.TheChickenbetaactincontainsaCpGislandthatcanhelptokeepthepromoteractiveforlongerinstableculturewhencomparedtotheCMVpromoter.Theubiquitinpromoterthatisdrivingexpressionofthepuromycingenedemonstratesahighlevelofexpressioninmostcelltypes.ItdemonstratessimiliarlevelsofexpressiontothecommonlyusedEF1-Alphapromoter.
SequenceandMap:
OtherInfo:
TranscriptionTermination:Thisplasmidcontainsthreealternativetranscriptionterminatorsformammalianbacterialandbacteriophage(T7)expression.Thismeansthatonlythepromoterneedstobechangedtoaltertheexpressionsystemyouareusing.Wesellmultiplepromotersthatcanbeusedineachofthesesystems.Thepresenceofeachterminatordoesnotreduceexpressioninthealternativesystems.
Cloning:
Cloninginagene:Thisplasmidcontainsagenewithinthemainmultiplecloningsite(NotI-ClaI).Anyplasmidthatwesellwherethegeneisthisconfigurationwillbelocatedintheexactsamepositioninrelationtothestartandstopcodonofthegene.TheonlyexceptionstothisrulearefusionsproteinswherethefusiongenemaybepositionedatthefrontorendoftheMCStoallowgenefusion.
Bypositioningallofourgenesinthesamelocationitallowsthemtobetransferredbetweenplasmidsusingthesamecloningmethodandrestrictionsitesregardlessoftheplasmidbeingusedfromourproductrange.InsertinganewgeneintothisplasmidshouldbeeasilypossIBLeusingarangeofstandardrestrictionenzymesitesthatflankthegenecurrentlyinthevector.
Multiplecloningsitenotes:InthemultiplecloningsitetherearetwoimportantrestrictionsitescalledBsgIandBseRIsites.ThesesitesbothcuttheDNAatthesamepositionandcleavethestopcodonofthegeneinthemultiplecloningsiteinthisplasmidtherebyproducingaTAoverhang.Thisoverhangiscompatiblewithanyofourpeptideorreporterfusiontagplasmidsalsocutwitheitheroftheseenzymes.ThisallowsseamlessC-terminalfusionstobemadewiththegeneinthismultiplecloningsiteusingasinglecloningstepfromourC-terminalpeptideandreportertagproductrange.NormallytheeasiestmethodistoclonetheC-terminaltagfromourotherplasmidproductsintothisplasmidusingBsgIorBseRIandthedownstreamClaIrestrictionsite.
BseRIandBsgIsitesarenon-palindromicandcleaveadefinednumberofbasesawayfromtheirbindingsites.Thisallowsthemtocuttheupstreamstopcodoninthegeneinthisplasmidregardlessofthegenesequence.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
