PlasmidInfo:
PlasmidInformation
ProductName:pSF-U6
ProductCode:OG51
Size(bp):3932bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:HumanU6promoter
PlasmidPurpose:
TheexpressionofshorthairpinRNAineukaryoticcells.TheU6istranscribedbyRNApolymeraseIII(RNApolIII)whichterminatestranscriptionatastringofthymidineresiduestypicallyaminimumoffiveresiduesinmammaliancells.UnlikemostSnapFastTMvectorsthisvectorcontainsanRNApolIIIterminatordownstreamoftheXhoIsitealthoughitismorecommontoinsertaterminatorsequenceattheendofthesequenceyouinsertintotheMCSthatencodesashRNA.
SequenceandMap:
OtherInfo:
TranscriptionTermination:ThisvectorcontainsanRNApolymeraseIIIpromoter(U6)whichwillterminateatastringofthymidineresidues.Typicallythisis5-6Tresiduesinlength.Thisplasmidcontainsastringofthymidinesinthemultiplecloningsitetoallowtermination.
Cloning:
Multiplecloningsitenotes:ThisvectorhasamodifiedMCScomparedtostandardplasmids.TheATGstartcodonintheNcoIsitehasbeenremovedashavetheShine-DalgarnoandKOZAKsequencesasthesearenotrequiredforRNAPol3expressedtranscripts.
TheterminatorforRNApolymeraseIIIhasbeenpositionedimmediatelydownstreamoftheXhoIsite.ThereisanadditionalstringofThymidineresidues(theterminatorsequenceforRNAPolIII)attheendoftheSV40polyAregionhoweverusingthissitewillproducealongtranscriptandhasnotbeenvalidated.WerecommendinsertingyourhairpinDNAsequencebetweenHind3andXhoI.ThetheoreticalRNAposition+1isthefirstbaseoftheNotIrestrictionsite.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
