>
产品中心 >
Sandwich_method_ELISA >
Oxford Genetics/pSF-OXB20-RrnB_(previously_pSF-RecA-RrnB) (OG179) RrnB Single Terminator Plasmid/OG179/1 Ea
PlasmidInfo:
PlasmidInformation
ProductName:pSF-OXB20-RrnB_(previously_pSF-RecA-RrnB)
ProductCode:OG179
Size(bp):3601bp
BacterialAntibioticSelection:KanR
OriginandCompatibility:pUChighcopyderivedfrompBR322
BacterialCopyNumber:500-700percell
Promoter:OXB20strongconstitutivebacterialpromoter
PlasmidPurpose:
Expressioninbacterialcells.TheRecApromoterisaverystrongconstitutivepromoter.Itisapproximately650xstrongerthantheconstitutiveAraBADpromoterthatwealsosell.TheRecApromoterisnormallyregulatedbytherepressorLexA.Inthispromoterthisbindingsitehasbeenablatedtoenableconstitutiveexpression.ThisplasmidisusedwhentheT7terminatorandtheSV40poly-adenylationsitesthatarefoundwithinmostofourvectorsarenotrequired.
PromoterExpressionLevel:Thisplasmidcontainsaconstitutivebacterialpromoterthatdoesnotrequireinduction.Itisthestrongestbacterialpromoterwesellandthiscancausesolubilityandexpressionproblemswithsomeproteins.WealsoofferarangeofotherbacterialpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.
SequenceandMap:
OtherInfo:
TranscriptionTermination:ThisplasmidcontainstheE.coliRrnBRhodepdendenttranscriptionalterminator.UnlikemostofourotherplasmidproductswhichcontainthreeterminatorsfordifferentorganismtypesthisvectoronlycontainstheRrnBterminatorregion.ThisplasmidisonlynormallyusedwhenthepresenceoftheothertwotranscriptionterminatorsnormallyinourvectorsisnotdesirableorspacerestrictionsinagivenBIOLOGicalsystemareimportant.
Cloning:
CloninginaGene:Thisplasmidhasbeendesignedtobecompatiblewitharangeofcloningtechniques.Themultiplecloningsitecontainsarangeofstandardcommonlyusedrestrictionsitesforcloning.UsingthesesitesgenescanbeinsertedusingstandardcloningmethodswithDNAligase.Othermethodssuchasligaseindependentcloning(LIC)GibsonAssemblyInFusionHDorSeamlessGeneArtcanalsobeusedandbecauseallofourplasmidsarebasedonthesamebackbonethesamemethodcanbeusedforcloningintoallofourcataloguevectors.
Multiplecloningsitenotes:ThereareafewimportantsiteswithintheMCS.TheseincludetheNcoIsitetheXbaIsiteandtheBsgIandBseRIsites.TheNcoIsitecontainsastartcodonthatisimmediatelydownstreamofbothaKozakandShine-Dalgarnoribosomalbindingsite.Theseallowforoptimalpositioningofgeneswhenthestartcodonisplacedinthislocation.IfthisisnotrequiredandyouwishtouseadownstreamsiteforgenecloningyoucanremovetheNcoIsitebycleavingtheplasmidwithKpnI.
TheXbaIsitecontainsastopcodon.ThisstopcodonispositionedinaspecificpositioninrelationtotheBsgIandBseRIsitesthatareimmediatelydownstream.WheneitherBseRIorBsgIcleavetheplasmidtheyproduceaTAoverhangfromthestopcodonintheXbaIsitethatiscompatiblewithallofourpeptidetagplasmidscutwiththesamesites.BseRIandBsgIsitesarenon-palindromicandcleaveadefinednumberofbasesawayfromtheirbindingsite.
WheneverwecloneageneintoourmultiplecloningsitewealwayspositionthestartandstopcodoninthesamepositionsintheMCS.IfthestartandendsofthegenesarenotcompatiblewithNcoIandXbaIweextendthesequencetothenearestexternalsitesbutkeepthestartandstopcodonslocationsconsistent.
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
