Description:

pOET2N/C_6xHisisabaculovirustransfervectordesignedforhighlevelexpressionofforeigngenesunderthepowerfulAcMNPVpolyhedron(polh)promoter.ThevectorencodesanNterminal6×His-Tag®fusionsequencethatmaybeutilizediftheinsertincludesastopcodon.Thisgreatlyeasesthepurificationoftherecombinantproteinsincethe6×His-containingfusionproteinsbindwithhighaffinitytoNi-NTAAgarose.Ifrequired,the6×His-Tag®canberemovedbyincubatingthefusionproteininthepresenceoftheproteinasecleavageenzymeThrombin.Thereisalsoa6×His-Tag®forC-terminalfusionswheretheinsertsownstartcodoncanbeusedtoreplacethestartcodonsuppliedinpOET2N/C_6xHis.pOET2N/C_6xHisissmallerthanotheravailabletransfervectors(4626bp)whichgreatlyfacilitatethecloningsteps.IthasaColE1originofreplicationandanampicillinresistancegeneforselectioninE.coli.Thepolhsequenceshavebeenreplacedbyamultiplecloningsite(MCS)containinguniquerestrictionsitesforinsertionoftheforeigngeneinthecorrectorientation,asshownonthecircularmap.ThecodingstrandoftheMCSastranscribedfromthepolhpromoterisshownbelowthecircularmap.ThePacIsiteattheendoftheMCSprovidestranslationalstopcodonsinallthreereADIngframesforexpressionoftruncatedproteins.TheAcMNPVsequencesflankingthegeneinthetransfervectorsMCSallowrecombinationwiththeviralDNAtoinserttheexpressioncassetteintothepolhlocus.pOET2N/C_6xHisiscompatIBLewithanybaculovirussystemthatutilizeshomologousrecombinationininsectcells.

AdditionalInformation:

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Name	pOET2N/C_6xHis™TransferVector
Concentration	LotSpecific
StABIlity	1year
Storage	-20C
IntendedUse	ResearchUseOnly