PlasmidInfo:
PlasmidInformation
ProductName:pSF-CMV-Ub_AscI
ProductCode:OG399
Size(bp):5595bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:Cytomegalovirus(CMV)immediateearlypromoter/HumanUbiquitinPromoter
ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.However,therearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.Forthisreason,westockarangeofotherpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.
{tab=PlasmidPurpose}
Purpose:ThisplasmidcontainstwomultiplecloningsitesthatallowtheexpressionoftwogenesinmammaliancellsonedrivenbytheCMVpromoterandonebythehumanUbiquitin(Ub)promoter.TheCMVpromoterissignificantlystrongerthantheUbpromoter(relativeexpressionwillvarywithcelltype)howevertheUbpromoterisreportedtobelesseffectedbymethylationthantheCMVpromoter.
Thisplasmidcontainstwomultiplecloningsitesandallowstheinsertionoftwogenes,onedownstreamoftheCMVpromoterandonedownstreamofthehumanUbiquitinpromoter.
SequenceandMap:
Cloning:
MainMultipleCloningSiteNotes:ThereisastartcodonintheNcoIsitethatcanberemovedbydigestionwithKpnIifrequired.TheMCSforgeneinsertionsextendsfromNotItoXbaI,buttheShine-DalgarnosequencesandKOZAKribosomalentrysitesequencesarealignedwiththestartcodonintheNcoIsite.
SecondMultipleCloningSiteNotes:IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
