PlasmidInfo:
PlasmidInformation
ProductName:pSF-H1
ProductCode:OG52
Size(bp):3801bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:HumanH1promoter
PlasmidPurpose:
TheexpressionofshorthairpinRNAineukaryoticcells.TheH1istranscribedbyRNApolymeraseIII(RNApolIII)whichterminatestranscriptionatastringofthymidineresiduestypicallyaminimumoffiveresiduesinmammaliancells.UnlikemostSnapFastTMvectorsthisplasmidcontainsanRNApolIIIterminatordownstreamoftheXhoIsitealthoughitismorecommontoinsertaterminatorsequenceattheendofthesequenceyouinsertintotheMCSthatencodesashRNA.
SequenceandMap:
OtherInfo:
TranscriptionTermination:ThisvectorcontainsanRNApolymeraseIIIpromoter(H1)whichwillterminateatastringofthymidineresidues.Typicallythisis5-6Tresiduesinlength.Thisplasmidcontainsastringofthymidinesinthemultiplecloningsitetoallowtermination.
Cloning:
Multiplecloningsitenotes:ThisvectorhasamodifiedMCScomparedtostandardplasmids.TheATGstartcodonintheNcoIsitehasbeenremovedashavetheShine-DalgarnoandKOZAKsequencesasthesearenotrequiredforRNAPol3expressedtranscripts.
TheterminatorforRNApolymeraseIIIhasbeenpositionedimmediatelydownstreamoftheXhoIsite.ThereisanadditionalstringofThymidineresidues(theterminatorsequenceforRNAPolIII)attheendoftheSV40polyAregionhoweverusingthissitewillproducealongtranscriptandhasnotbeenvalidated.WerecommendinsertingyourhairpinDNAsequencebetweenHind3andXhoI.ThetheoreticalRNAposition+1isthefirstbaseoftheNotIrestrictionsite.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
