Description:

pOET1N_6xHisisabaculovirustransfervectordesignedforhighlevelexpressionofforeigngenesunderthepowerfulAcMNPVpolyhedron(polh)promoter.ThevectorencodesanoptionalN-terminal6×His-Tagfusionsequencethatmaybeutilizediftheinsertallowsread-throughinthecorrectreADIngframe.Thisgreatlyeasesthepurificationoftherecombinantproteinsincethe6×His-containingfusionproteinsbindwithhighaffinitytoNi-NTAAgarose.Ifrequired,the6×His-TagcanberemovedbyincubatingthefusionproteininthepresenceoftheproteinasecleavageenzymeThrombin.pOET1N_6xHisissmallerthanotheravailabletransfervectors(4598bp)whichgreatlyfacilitatethecloningsteps.IthasaColE1originofreplicationandanampicillinresistancegeneforselectioninE.coli.Thepolhsequenceshavebeenreplacedbyamultiplecloningsite(MCS)containinguniquerestrictionsitesforinsertionoftheforeigngeneinthecorrectorientation,asshownonthecircularmap.ThecodingstrandoftheMCSastranscribedfromthepolhpromoterisshownbelowthecircularmap.ThePacIsiteattheendoftheMCSprovidestranslationalstopcodonsinallthreereadingframesforexpressionoftruncatedproteins.TheAcMNPVsequencesflankingthegeneinthetransfervectorsMCSallowrecombinationwiththeviralDNAtoinserttheexpressioncassetteintothepolhlocus.pOET1N_6xHisiscompatIBLewithanybaculovirussystemthatutilizeshomologousrecombinationininsectcells. 

AdditionalInformation:

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Name	pOET1N_6xHistransferplasmid
Concentration	LotSpecific
StABIlity	6months
Storage	-20
IntendedUse	ResearchUseOnly