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Oxford Genetics/pSF-CMV-COOH-P2A-3 (OG65) FMDV P2A Plasmid/OG65/1 Ea
PlasmidInfo:
PlasmidInformation
ProductName:pSF-CMV-COOH-P2A-3
ProductCode:OG65
Size(bp):4280bp
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
