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当前位置: 首页 > 产品中心 > vector > 牛津遗传学/pSF-CMV-Puro-COOH-3C-Strep-6His(OG1256)C-末端6 His和Strep双标签哺乳动物质粒/OG1256/1
商品详细牛津遗传学/pSF-CMV-Puro-COOH-3C-Strep-6His(OG1256)C-末端6 His和Strep双标签哺乳动物质粒/OG1256/1
牛津遗传学/pSF-CMV-Puro-COOH-3C-Strep-6His(OG1256)C-末端6 His和Strep双标签哺乳动物质粒/OG1256/1
牛津遗传学/pSF-CMV-Puro-COOH-3C-Strep-6His(OG1256)C-末端6 His和Strep双标签哺乳动物质粒/OG1256/1
商品编号: OG1256
品牌: oxgene
市场价: ¥5321.60
美元价: 3192.96
产地: 美国(厂家直采)
公司:
产品分类: 载体
公司分类: vector
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍

PlasmidInfo:

PlasmidInformation

ProductName:pSF-CMV-Puro-COOH-3C-Strep-6His

ProductCode:OG1256

Size(bp):6216bp

BacterialAntibioticSelection:KanR

OriginandCompatibility:pUChighcopyderivedfrompBR322

BacterialCopyNumber:500-700percell

Promoter:Cytomegalovirus(CMV)immediateearlypromoter/HumanUbiquitinPromoter

PlasmidPurpose:

Thisplasmidisdesignedtoexpresstaggedproteinsinmammaliancellseitherbytransienttransfectionorbycreatingstablecelllines.ItcontainsapuromycinresistanceexpressioncassetteusingthehumanUbiquitinpromotertodriveexpressionandallowfortheselectionofcellscontainingtheplasmid.

AbouttheCleavageTag:

Thisplasmidalsoencodesaproteasecleavagesitethatisdesignedtobepositionedbetweenyourgeneofinterestandthetagtoallowtheremovalofthetagfollowingproteinpurificationorisolation.Thisplasmidcontainsa3Ccleavagetag.Theproteinsequenceofthecleavagetagis:LEVLFQ?GP.HumanRhinovirus(HRV)3CProteaseisahighlyspecificproteasethatcleavesbetweentheGluandGlyresiduesofitsrecognitionsite.Itisoftenproducedwiththetrademname"PreScissionprotease".

Formoreinformationonwhichcleavagetagtouseseeourcleavagetagguide.

PromoterExpressionLevel:

ThisplasmidcontainsthemammalianCMVpromotertodrivegeneexpression.WehavetestedallofourmammalianpromotersinarangeofcelltypesandCMVisconsistentlythestrongestinthosewehavestudied.HowevertherearemanyreportsoftheCMVpromoterdemonstratingsilencingbymethylationinlong-termculture.ForthisreasonwestockarangeofotherpromotersthatarecompatIBLewiththisplasmidandareavailableonrequest.

AboutthePeptideTag:

Thisplasmidcontainsac-terminalStrepaffinitytagthatcanbefusedtoageneofinteresttoallowproteindetectionand/orpurification.Thesequenceofthetagis:WSHPQFEK

Formoreinformationonthemethodsthatcanbeusedtopurifyproteinspleaseseeourproteintagguide.

ThisplasmidalsocontainsasecondaryHexa-Histidine(6His)tagproteintag.Thesequenceofthistagis:HHHHHH

Weprovidearangeofdualpeptidetagplasmids.ThisisbecausesomepeptidetagsprovidespecificBIOLOGicalproperties(e.gsmallmoleculeaffinitynewepitopessolubilityorproteinsecretion)thatarenotprovidedbyothers.

SequenceandMap:

OtherInfo:

TranscriptionTermination:

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

Cloning:

MakingProteinFusions:

ThisplasmidhasbeendesignedtoallowthreetypesofcloningintothemainMCStojoinacodingsequencewiththetag.

SnapFusionCloning:

IfyouwouldliketofuseyourcodingsequencetothetagwithminimaladditionalbasesyoucanuseourSnapFusiontechnology.ThisprocessinvolvesamplifyingyourgenebyPCRtoaddspecificrestrictionsitesontotheends.WhenthesesitesarecuttheyproduceanoverhangthatiscompatiblewiththisplasmidcutwithBseRIorBsgI.

Toinsertyourgene:


1:Amplifyyourgenewithprimersdesignedusingthisspreadsheet
2:CuttheplasmidwitheitherBseRIorBsgI.*
3:Cutyourgenewiththeenzymeyouaddedusingthespreadsheet(anyofAcuIBpmIBpuEIBseRIBsgIEciI).
4:ClonethegeneintotheplasmidusingDNAligase.

UsingthismethodwithanN-terminaltagplasmidwillresultinthetagcodingsequenceimmediatelyfollowedbyyourgenesATGstartcodonatthejoin.Thisresultsinaseamlessfusionofthetwosequenceswithnoextrabasesbeingadded.UsingthismethodonC-terminaltagplasmidswillconvertyourgenesstopcodonintoaTAC(TyrY)codonfollowedbytheplasmidtagcodingsequence.Thisresultsinnoextrabasesbetweenyourgeneandthetag.Seethediagrambelowformoreinformation.

*PleasenotethatinsectexpressionplasmidscannotbecutwithBsgIonlyBseRIbecauseofunavoidableconflictingsitesinthebackbone.AlsoYeastplasmidscanonlybecutwithBsgInotBseRIbecauseofconflictingsitesinthebackbone.

Usingthistechniquewillcreateagenefragmentthatcanbeligatedintoanyorour>1500peptideandreportertagplasmids.Ifyouuseoneoftheothertechniquesbelow(GibsonInFusionSeamlessorLIC)youwillneednewprimersforeveryvectoryoucloneintobecausethearmsofhomologywillchangeaccordingtothetagplasmidyouarecloninginto.

Ifyoufindthatyourgenesequencehassitesinitthatmakeusingthiscloningstrategydifficultyoucanstilluseoneofthealternativemethodsbelow(e.g.standardcloningorGibsoncloning).

OpenthePrimerDesignTooltohelpyoudesignprimersforcloningyourgeneinourSnapFusiontechnique.

StandardEnzymes:

Ifyouarenotconcernedaboutleavingafewextrabasesbetweenthetagcodingsequenceandyourgeneyoucancloneyourgeneintothevectorusingstandardcloningrestrictionenzymes.Thisstrategywillrequireyoutochoosewhichenzymesyouwanttousetocloneyourgene.

OpenthePrimerDesignToolwhichprovidesprimerswithdifferentenzymechoicespositioningyourgeneasclosetothetagaspossibleineachcase.Pleasenotethatstandardenzymeswillalwaysleaveadditionalnucleotidesbetweenyourgeneandthetagbutusingthespreadsheetwillensurethetagandgeneareinframe.

Gibsoncloning/InfusionHD/GeneArtSeamless/LigaseIndependentCloning(LIC)Methods:

ThesecloningtechniquesusereagentssoldbyothercompaniesandallowyoutofusesequencestogetherusingenzymesthatchewbacktheDNAtoleaveoverlappingends/overhangs.ThesubsequentmethodofjoiningtheDNAdependsonthekitused.Touseoneofthesetechniquesyoucaneitherdesignyourownprimersoryoucanusethespreadsheetbelowtohelpwiththedesign.

OpenthePrimerDesignTooltohelpyoudesignprimersforcloningyourgeneusingGibsonassemblyInfusionHDGeneArtSeamlesscloningorLigaseIndependentCloning(LIC)techniques.

IPStatus:

IntellectualPropertyStatus

ThisproductispartofourSnapFastplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere

品牌介绍
OXGENE的技术平台和专家解决方案加速了细胞和基因疗法的发现,开发和制造。我们解决了现代生物学中基因治疗,抗体治疗和基因编辑方面最重要和最具挑战性的问题。我们的技术可以实现精确而强大的哺乳动物细胞工程。我们以自动化和信息学为导向的方法意味着我们解决了其他人无法推进新疗法交付的问题。