Oxford Genetics/pSFCMVFLucPGKCas9WTBgH_Pac1 (OG3531) CRISPR/Cas9 Expression Plasmid/OG3531/1 Ea

PlasmidInfo:

PlasmidInformation

ProductName:pSF-CMV-FLuc-PGK-Cas9WT-BgH_Pac1

ProductCode:OG3531

Size(bp):10841bp

BacterialAntibioticSelection:AmpR

OriginandCompatibility:pUChighcopyderivedfrompBR322

BacterialCopyNumber:500-700percell

Promoter:Cytomegalovirus(CMV)immediateearlypromoter/MousePhosphoglycerateKinase(PGK)Promoter

PlasmidPurpose:

ThisdualexpressionvectorisdesignedtoexpressCas9fromthePGKpromoterinthePac1siteaswellasthefireflyluciferasereportergenefromaCMVpromoterinthemainMCS.

PromoterExpressionLevel:

ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.HowevertherearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.Forthisreasonwestockarangeofotherpromotersthatarecompatiblewiththisplasmidandareavailableonrequest.

SequenceandMap:

OtherInfo:

TranscriptionTermination:

Thisplasmidcontainsthreealternativetranscriptionterminatorsformammalianbacterialandbacteriophage(T7)expression.Thismeansthatonlythepromoterneedstobechangedtoaltertheexpressionsystemyouareusing.Wesellmultiplepromotersthatcanbeusedineachofthesesystems.Thepresenceofeachterminatordoesnotreduceexpressioninthealternativesystems.TheCas9proteinisterminatedbytheBovinegrowthhormonepolyAsignal.

Cloning:

Cloninginagene:

Thisplasmidcontainsagene(FLuc)withinthemainmultiplecloningsite(NotI-ClaI).Anyplasmidthatwesellwherethegeneisthisconfigurationwillbelocatedintheexactsamepositioninrelationtothestartandstopcodonofthegeneintheplasmid.TheonlyexceptionstothisrulearefusionsproteinswherethefusiongenemaybepositionedatthefrontorendoftheMCStoallowgenefusion.

Bypositioningallofourgenesinthesamelocationitallowsthemtobetransferredbetweenplasmidsusingthesamecloningmethodandrestrictionsitesregardlessoftheplasmidbeingusedfromourproductrange.Insertinganewgeneintothisplasmidshouldbeeasilypossibleusingarangeofstandardrestrictionenzymesitesthatflankthegenecurrentlyinthevector.

Multiplecloningsitenotes:

ThereareafewimportantsiteswithintheMCS.TheseincludetheNcoIsitetheXbaIsiteandtheBsgIandBseRIsites.TheNcoIsitecontainsastartcodonthatisimmediatelydownstreamofbothaKozakandShine-Dalgarnoribosomalbindingsite.Theseallowforoptimalpositioningofgeneswhenthestartcodonisplacedinthislocation.IfthisisnotrequiredandyouwishtouseadownstreamsiteforgenecloningyoucanremovetheNcoIsitebycleavingtheplasmidwithKpnI.

BseRIandBsgIsitesarenon-palindromicandcleaveadefinednumberofbasesawayfromtheirbindingsites.Thisallowsthemtocuttheupstreamstopcodoninthegeneinthisplasmidregardlessofthegenesequence.

IPStatus:

IntellectualPropertyStatus

ThisproductispartofourSnapFastProplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere

对照品标准品，生物试剂盒，gibco胎牛血清，试剂采购网，中国试剂网，细胞因子试剂盒