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Oxford Genetics/pSF-CMV-NH2-V5-EKT1 (OG327) N-Terminal V5 Tag Plasmid/OG327/1 Ea188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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Oxford Genetics/pSF-CMV-NH2-V5-EKT1 (OG327) N-Terminal V5 Tag Plasmid/OG327/1 Ea

PlasmidInfo:

PlasmidInformation

ProductName:pSF-CMV-NH2-V5-EKT1

ProductCode:OG327

Size(bp):4313bp

BacterialAntibioticSelection:KanR

OriginandCompatibility:pUChighcopyderivedfrompBR322

BacterialCopyNumber:500-700percell

Promoter:Cytomegalovirus(CMV)immediateearlypromoter

PlasmidPurpose:

ThisvectoraddsaV5epitopetagtotheN-terminusofaproteinthatisencodedwithinthemultiplecloningsite.ThistagallowsthedetectionandpurificationofataggedproteinusingantibodiesraisedagainstV5.TheV5tagcodingsequenceisGKPIPNPLLGLDST.Thereisanenterokinase(EKT)cleavagesite(DDDDK)immediatelydownstreamoftheV5tagthatcanbeusedtoremovetheV5tagfromapurifiedprotein.Itcleavesafterthelysineresidue.

PromoterExpressionLevel:

ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.HowevertherearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.ForthisreasonwestockarangeofotherpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.

SequenceandMap:

OtherInfo:

TranscriptionTermination:

Thisplasmidcontainsthreealternativetranscriptionterminatorsformammalianbacterialandbacteriophage(T7)expression.Thismeansthatonlythepromoterneedstobechangedtoaltertheexpressionsystemyouareusing.Wesellmultiplepromotersthatcanbeusedineachofthesesystems.Thepresenceofeachterminatordoesnotreduceexpressioninthealternativesystems.

Cloning:

CloninginaGene:

Thisvectorhasbeendesignedtoallowtheadditionofapeptidetagtotheendofaproteinofinterestusingstandardcloningtechniques.

MultipleCloningSiteNotes:

ThereisastartcodonintheNcoIsitecanberemovedbydigestionwithKpnIifrequired.TheMCSforgeneinsertionsextendsfromNotItoXbaIhoweverthetagresidesbetweentheNotIandHindIIIsites.ThereareShine-DalgarnosequencesandKOZAKsequencesalignedwiththestartcodonofthepeptidetag.

TheClaItoNheIsiteshaveotherfunctionssuchasaddingC-terminalpeptidetagssecondpromotersorIRESexpressioncomponents.TheBsgIandBseRIrestrictionsitescleavewithinthestopcodonintheXbaIsiteandallowtheretrospectivefusionofC-terminalpeptidetagssequencesifthestopcodonisplacedinthisposition.

IPStatus:

IntellectualPropertyStatus

ThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere

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