pSecTag2A哺乳动物表达载体说明

pSecTag2A编号  载体名称  北京华越洋生物VECT6096  pSecTag2  A  pSecTag2A载体基本信息载体名称:pSecTag2A质粒类型:哺乳细胞表达载体；分泌表达载体高拷贝/低拷贝:高拷贝启动子:CMV克隆方法:多克隆位点，限制性内切酶载体大小:5159bp5'测序引物及序列:T7-F：5'd[TAATACGACTCACTATAGGG]3'3'测序引物及序列:--载体标签:6XHis,myc载体抗性:Ampicillin筛选标记:Zeocin备注:含有前导序列IgK，引导重组蛋白分泌表达。稳定性:瞬表达组成型:组成型病毒/非病毒:非病毒pSecTag2A载体质粒图谱和多克隆位点信息pSecTag2A载体简介pSecTag2andpSecTag2/Hygroaremammalianexpressionvectorsdesignedforthesecretion,purification,anddetectionoffusionproteins.EachvectorhasalargemultiplecloningsiteinthreereadingframestosimplifycloninginframewiththeN-terminalsecretionsignal.Thevectors(Figure1)offerthefollowingfeatures:SecretionsignalfromtheV-J2-CregionofthemouseIgkappa-chainforefficientsecretionofrecombinantproteins(Figure2)Cytomegalovirus(CMV)promoterforhigh-levelconstitutiveexpressionC-terminalpolyhistidine(6xHis)tagforrapidpurificationwithnickel-chelatingresinanddetectionwithanAnti-His(C-term)AntibodyC-terminalc-mycepitopefordetectionwithanAnti-mycAntibodyBovineGrowthHormone(BGH)polyadenylationsignalandtranscriptionterminationsequencetoenhancemRNAstabilitySV40originforepisomalreplicationandsimplevectorrescueincelllinesexpressingthelargeTantigen(e.g.,COS-1,COS-7)ThepSecTag2vectorscarrytheZeocinresistancegeneforcost-effectiveselectioninmammaliancells.ZeocinselectioncanalsobeusedinE.coli.ThepSecTag2/HygrovectorshavetheHygromycinBresistancegeneforselectionofstablemammaliancelllines.引用及参考文献Moleculardissectionofthealpha-dystroglycan-andintegrin-bindingsiteswithintheglobulardomainofhumanlaminin-10.IdoH,HaradaK,FutakiS,HayashiY,NishiuchiR,NatsukaY,LiS,WadaY,CombsAC,ErvastiJM,SekiguchiK,JBiolChem(2004)279:10946-10954Productusage:The5cDNAwasinsertedintotheNheI/PmeIsitesofpcDNA3.1.Forsite-directedmutagenesisoftheLG3module,thecDNAfragmentencodingLG3wasexcisedfrompcDNA-5LG4–5withAscIandPmeIandreclonedintopSecTag2AattheAscI/PmeIsites.RecombinantproteinwasproducedusiProhaptoglobinisproteolyticallycleavedintheendoplasmicreticulumbythecomplementC1r-likeprotein.WicherKB,FriesE,ProcNatlAcadSciUSA(2004)101:14390-14395Productusage:HEK293andCOS-1cellsweretransientlytransfectedwithC1r-LPcDNAclonedintopcDNA6-myc-hisorwithproHpcDNAclonedintopSecTag.After24h,thecellswererinsedtwIcewithPBSandgrownfor48hinaserum-freemedium.Thecellmediawerethencollected.pSecTag2A载体序列ORIGIN1GACGGATCGGGAGATCTCCCGATCCCCTATGGTCGACTCTCAGTACAATCTGCTCTGATG61CCGCATAGTTAAGCCAGTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCG121CGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGC181TTAGGGTTAGGCGTTTTGCGCTGCTTCGCGATGTACGGGCCAGATATACGCGTTGACATT241GATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATA301TGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACC361CCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCC421ATTGACGTCAATGGGTGGACTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGT481ATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATT541ATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCA601TCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTG661ACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACC721AAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCG781GTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCA841CTGCTTACTGGCTTATCGAAATTAATACGACTCACTATAGGGAGACCCAAGCTGGCTAGC901CACCATGGAGACAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCAC961TGGTGACGCGGCCCAGCCGGCCAGGCGCGCCGTACGAAGCTTGGTACCGAGCTCGGATCC1021ACTCCAGTGTGGTGGAATTCTGCAGATATCCAGCACAGTGGCGGCCGCTCGAGGAGGGCC1081CGAACAAAAACTCATCTCAGAAGAGGATCTGAATAGCGCCGTCGACCATCATCATCATCA1141TCATTGAGTTTAAACCCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGT1201TGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTC1261CTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGG1321TGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGA1381TGCGGTGGGCTCTATGGCTTCTGAGGCGGAAAGAACCAGCTGGGGCTCTAGGGGGTATCC1441CCACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGAC1501CGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGC1561CACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGCATCCCTTTAGGGTTCCGATT1621TAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGG1681GCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAG1741TGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTT1801ATAAGGGATTTTGGGGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATT1861TAACGCGAATTAATTCTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCC1921CCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCAGGTGTGGAAAG1981TCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACC2041ATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCT2101CCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCTGCCTCT2161GAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAGCTC2221CCGGGAGCTTGTATATCCATTTTCGGATCTGATCAGCACGTGTTGACAATTAATCATCGG2281CATAGTATATCGGCATAGTATAATACGACAAGGTGAGGAACTAAACCATGGCCAAGTTGA2341CCAGTGCCGTTCCGGTGCTCACCGCGCGCGACGTCGCCGGAGCGGTCGAGTTCTGGACCG2401ACCGGCTCGGGTTCTCCCGGGACTTCGTGGAGGACGACTTCGCCGGTGTGGTCCGGGACG2461ACGTGACCCTGTTCATCAGCGCGGTCCAGGACCAGGTGGTGCCGGACAACACCCTGGCCT2521GGGTGTGGGTGCGCGGCCTGGACGAGCTGTACGCCGAGTGGTCGGAGGTCGTGTCCACGA2581A