PlasmidInfo:
PlasmidInformation
ProductName:pSF-CMV-COOH-P2A-2
ProductCode:OG64
Size(bp):4279bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:Cytomegalovirus(CMV)immediateearlypromoter
PlasmidPurpose:
ThisvectoraddsaFootandMouthDiseaseP2ApeptidebetweentheprimaryMCS(NotI-XbaIbutXbaIisablatedinthisvector)andthefusionMCS(ClaI-NheI).TheP2Apeptidecodingsequenceis:GSGATNFSLLKQAGDVEENPGP.Thepeptideself-cleavesbetweenthepenultimateglycineresidueandthefinalprolineresidue.Thisleavesthefirst(upstream)proteininthepolypeptidewiththemajorityoftheaminoacidsoftheP2ApeptideontheC-terminuswhilstthelast(downstream)proteinhasaprolineremainingontheN-terminus.Wehavenotconfirmedifsecretoryproteinsareabletodetachfromcytosolicproteinswhenfusedusingthismethod.Theintracellulartraffickingofyourproteinsshouldthereforebeconsidered.
PromoterExpressionLevel:ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.HowevertherearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.ForthisreasonwestockarangeofotherpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.
SequenceandMap:
OtherInfo:
TranscriptionTermination:Thisplasmidcontainsthreealternativetranscriptionterminatorsformammalianbacterialandbacteriophage(T7)expression.Thismeansthatonlythepromoterneedstobechangedtoaltertheexpressionsystemyouareusing.Wesellmultiplepromotersthatcanbeusedineachofthesesystems.Thepresenceofeachterminatordoesnotreduceexpressioninthealternativesystems.
Cloning:
MultipleCloningSiteNotes:ThereisastartcodonintheNcoIsitecanberemovedbydigestionwithKpnIifrequired.TheMCSforgeneinsertionsinmostSnapFastvectorsextendsfromNotItoXbaI.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
