pcDNA5/FRT/TO载体质粒图谱、序列 - 生物风
文章推薦指數: 80 %
pcDNA5/FRT/TO 载体价格6500元,pcDNA5/FRT/TO 载体质粒图谱(Vector map),载体序列(Sequence)见下文,载体质粒抗性为氨苄青霉素,质粒大小为5137bp,测序引物为CMV ...
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热点推荐胎牛血清蛋白激酶生物风国际试剂进出口物流中心热点推荐Gibco胎牛血清HyClone胎牛血清Sigma胎牛血清Corning胎牛血清BI胎牛血清Gemini胎牛血清PAN胎牛血清Bovogen胎牛血清Hyclone澳洲胎牛血清Sigma澳洲小牛血清HyClone细胞培养基Gibco细胞培养基ATCC细胞培养基蛋白激酶EMSF930无血清昆虫细胞培养基生物资源载体质粒菌种库细胞库基因cDNAAdd基因感受态生物资源ATCC菌种库ATCC细胞库普通细胞株其他菌种基因cDNA质粒表达质粒Add基因质粒和载体感受态细胞大肠杆菌表达载体哺乳细胞表达载体酵母表达载体昆虫表达载体腺病毒载体慢病毒载体逆病毒载体Gateway载体植物表达载体Crispr/Cas9载体广宿主载体无细胞表达载体其他类型载体细胞培养细胞培养基血清细胞添加剂转染试剂细胞培养Gibco血清Hyclone血清Sigma血清Corning血清PAN血清BI血清Gemini血清Bovogen血清Gibco培养基HyClone培养基ATCC细胞培养基昆虫细胞培养基细胞培养添加剂DMEMRPMI-1640DMEM/F12MEMEMEMLeibovitzL-15199培养基McCoy培养基双抗/三抗PBS细胞培养级水GibcoSF900II/III昆虫培养基EMSF930昆虫无血清培养基微生物与植物培养
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pcDNA5/FRT/TO
价格:6500元
联系方式:I47-825O-882O
相关技术服务:质粒构建 基因合成 质粒大提
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载体基本信息
出品公司:
Invitrogen
载体名称:
pcDNA5/FRT/TO
质粒类型:
哺乳动物细胞表达载体;cDNA表达载体;定向重组载体;四环素调控载体
高拷贝/低拷贝:
高拷贝
克隆方法:
限制性内切酶,多克隆位点
启动子:
CMVTO(CMV/TetO2)
载体大小:
5137bp
5'测序引物及序列:
CMVForward:5’-CGCAAATGGGCGGTAGGCGTG-3’
3'测序引物及序列:
BGHReverse:5’-TAGAAGGCACAGTCGAGG-3’
载体标签:
无标签
载体抗性:
氨苄青霉素
筛选标记:
潮霉素(Hygromycin)
克隆菌株:
TOP10,DH5α,JM109
宿主细胞(系):
Invitrogen公司出品的Flp-In细胞系,293、CV-1、CHO等
备注:
pcDNA5/FRT/TO载体是cDNA的表达与克隆载体;
CMVTO启动子受四环素调控;
含有FRT位点(重组酶识别位点),与pOG44载体共转染,利用重组酶进行定向重组。
产品目录号:
V6520-20
稳定性:
瞬表达或稳表达
组成型/诱导型:
诱导型
病毒/非病毒:
非病毒
载体质粒图谱和多克隆位点信息
载体简介
简介
pcDNAFRTTOvectorisa5.1kbinducibleexpressionvectordesignedforusewiththeFlp-InT-RExSystem.Thecontrolplasmid,pcDNA5FRTTOCAT,isalsoincludedforuseasapositivecontrolfortransfectionandexpressioninyourFlp-InT-RExhostcelllineofchoice.
pcDNA5/FRT/TOisa5.1kbinducibleexpressionvectordesignedforusewiththeFlp-InT-RExSystem(Cat.no.K6500-01)availablefromInvitrogen.WhencotransfectedwiththepOG44FlprecombinaseexpressionplasmidintoaFlp-InT-RExmammalianhostcellline,thepcDNA5/FRT/TOvectorcontainingthegeneofinterestisintegratedinaFlprecombinase-dependentmannerintothegenome.Expressionofthegeneofinterestmaybeinducedbytheadditionoftetracyclinetotheculturemedium.
pcDNA5/FRT/TO载体含有以下元件:
Ahybridhumancytomegalovirus(CMV)/TetO2promoterforhigh-level,tetracycline-regulatedexpressionofthegeneofinterestinawiderangeofmammaliancells(Anderssonetal.,1989;Boshartetal.,1985;HillenandBerens,1994;Hillenetal.,1983;Nelsonetal.,1987)
Multiplecloningsitewith10uniquerestrictionsitestofacilitatecloningthegeneofinterest
FLPRecombinationTarget(FRT)siteforFlprecombinase-mediatedintegrationofthevectorintotheFlp-InT-RExhostcellline
Hygromycinresistancegeneforselectionofstablecelllines(GritzandDavies,1983)
Thecontrolplasmid,pcDNA5/FRT/TO/CAT,isincludedforuseasapositivecontrolfortransfectionandexpressionintheFlp-InT-RExhostcelllineofchoice.
FormoreinformationabouttheFlp-InT-RExSystem,thepOG44plasmid,andgenerationoftheFlp-InT-RExhostcellline,refertotheFlp-InT-RExCoreKitmanual.
杂交CMV/TetO2启动子
ExpressionofyourgeneofinterestfrompcDNA5/FRT/TOiscontrolledbythestrongCMVimmediateearlyenhancer/promoter(Anderssonetal.,1989;Boshartetal.,1985;Nelsonetal.,1987)intowhich2copiesofthetetoperator2(TetO2)sequencehavebeeninsertedintandem.InsertionoftheseTetO2sequencesintotheCMVpromoterconfersregulationbytetracyclinetothepromoter.TheTetO2sequencesconsistof2copiesofthe19nucleotidesequence,5′-TCCCTATCAGTGATAGAGA-3′separatedbya2basepairspacer(HillenandBerens,1994;Hillenetal.,1983).Each19nucleotideTetO2sequenceservesasthebindingsitefor2moleculesoftheTetrepressor.FormoreinformationaboutthemechanismoftetracyclineregulationintheFlp-InT-RExSystem,refertotheFlp-InT-RExCoreKitmanual.
关于pcDNA5/FRT/TO需要注意的事项
ThepcDNA5/FRT/TOvectorcontainsasingleFRTsiteimmediatelyupstreamofthehygromycinresistancegeneforFlprecombinase-mediatedintegrationandselectionofthepcDNA5/FRT/TOplasmidfollowingcotransfectionofthevector(withpOG44)intoFlp-InT-RExmammalianhostcells.TheFRTsiteservesasboththerecognitionandcleavagesitefortheFlprecombinaseandallowsrecombinationtooccurimmediatelyadjacenttothehygromycinresistancegene.TheFlprecombinaseisexpressedfromthepOG44plasmid.
ThehygromycinresistancegeneinpcDNA5/FRT/TOlacksapromoterandanATGinitiationcodon;therefore,transfectionofthepcDNA5/FRT/TOplasmidaloneintomammalianhostcellswillnotconferhygromycinresistancetothecells.TheSV40promoterandATGinitiationcodonrequiredforexpressionofthehygromycinresistancegeneareintegratedintothegenome(intheFlp-InT-RExhostcellline)andareonlybroughtintothecorrectproximityandframewiththehygromycinresistancegenethroughFlprecombinase-mediatedintegrationofpcDNA5/FRT/TOattheFRTsite.
Flp重组酶介导的DNA重组
IntheFlp-InT-RExSystem,integrationofyourpcDNA5/FRT/TOinducibleexpressionconstructintothegenomeoccursviaFlprecombinase-mediatedintermolecularDNArecombination.ThehallmarksofFlp-mediatedrecombinationarelistedbelow.
RecombinationoccursbetweenspecificFRTsites(seebelow)ontheinteractingDNAmolecules
RecombinationisconservativeandrequiresnoDNAsynthesis;theFRTsitesarepreservedfollowingrecombinationandthereisminimalopportunityforintroductionofmutationsattherecombinationsite
Strandexchangerequiresonlythesmall34bpminimalFRTsite
FRT位点
TheFRTsite,originallyisolatedfromSaccharomycescerevisiae,servesasabindingsiteforFlprecombinaseandhasbeenwell-characterized(GronostajskiandSadowski,1985;Jayaram,1985;Sauer,1994;Senecoffetal.,1985).TheminimalFRTsiteconsistsofa34bpsequencecontainingtwo13bpimperfectinvertedrepeatsseparatedbyan8bpspacerthatincludesanXbaIrestrictionsite.Anadditional13bprepeatisfoundinmostFRTsites,butisnotrequiredforcleavage(Andrewsetal.,1985).WhileFlprecombinasebindstoallthreeofthe13bprepeats,strandcleavageactuallyoccursattheboundariesofthe8bpspacerregion
pcDNA5/FRT/TO载体使用流程如下:
载体序列
pcDNA5/FRT/TO5137bp
GACGGATCGGGAGATCTCCCGATCCCCTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTT
AAGCCAGTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACA
ACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCG
ATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTC
ATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCG
CCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCC
ATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCC
AAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTA
TGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGC
AGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAA
TGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACG
CAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTCCCTATCAGTGATAGAGATC
TCCCTATCAGTGATAGAGATCGTCGACGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCC
ACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTCCGGACTCTAGCGTTTAAACTTAAG
CTTGGTACCGAGCTCGGATCCACTAGTCCAGTGTGGTGGAATTCTGCAGATATCCAGCACAGTGGCGGCC
GCTCGAGTCTAGAGGGCCCGTTTAAACCCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCT
GTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAA
ATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAG
CAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCTTCTGAGGCG
GAAAGAACCAGCTGGGGCTCTAGGGGGTATCCCCACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTG
TGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCC
TTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGA
TTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTACCTAGAAGTTCC
TATTCCGAAGTTCCTATTCTCTAGAAAGTATAGGAACTTCCTTGGCCAAAAAGCCTGAACTCACCGCGAC
GTCTGTCGAGAAGTTTCTGATCGAAAAGTTCGACAGCGTCTCCGACCTGATGCAGCTCTCGGAGGGCGAA
GAATCTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATATGTCCTGCGGGTAAATAGCTGCGCCGATG
GTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCCGCGCTCCCGATTCCGGAAGTGCTTGA
CATTGGGGAATTCAGCGAGAGCCTGACCTATTGCATCTCCCGCCGTGCACAGGGTGTCACGTTGCAAGAC
CTGCCTGAAACCGAACTGCCCGCTGTTCTGCAGCCGGTCGCGGAGGCCATGGATGCGATCGCTGCGGCCG
ATCTTAGCCAGACGAGCGGGTTCGGCCCATTCGGACCGCAAGGAATCGGTCAATACACTACATGGCGTGA
TTTCATATGCGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCGTCAGTGCG
TCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGAAGTCCGGCACCTCGTGC
ACGCGGATTTCGGCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTGACTGGAGCGA
GGCGATGTTCGGGGATTCCCAATACGAGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGTATG
GAGCAGCAGACGCGCTACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCCGGGCGTATA
TGCTCCGCATTGGTCTTGACCAACTCTATCAGAGCTTGGTTGACGGCAATTTCGATGATGCAGCTTGGGC
GCAGGGTCGATGCGACGCAATCGTCCGATCCGGAGCCGGGACTGTCGGGCGTACACAAATCGCCCGCAGA
AGCGCGGCCGTCTGGACCGATGGCTGTGTAGAAGTACTCGCCGATAGTGGAAACCGACGCCCCAGCACTC
GTCCGAGGGCAAAGGAATAGCACGTACTACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGG
CTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTC
GCCCACCCCAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAA
ATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTG
TATACCGTCGACCTCTAGCTAGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTAT
CCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGA
GCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCA
TTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACT
GACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTAT
CCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAA
AAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCA
AGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGC
GCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCT
TTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCAC
GAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGAC
ACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTAC
AGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTG
AAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTG
GTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTC
TACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGG
ATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTT
GGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCAT
AGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCA
ATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCG
AGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGT
AAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCG
TCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGT
GCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACT
CATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGT
GAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATAC
GGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAA
ACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCA
GCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAA
TAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGG
TTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACA
TTTCCCCGAAAAGTGCCACCTGACGTC
载体质粒图谱pdf版和相关资料下载
载体应用举例
pcDNA5/FRT/TO-TOPO2015-04-26
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- 4pcDNA 5/FRT/TO - Thermo Fisher Scientific
T-REx™ mammalian host cell line, the pcDNA™5/FRT/TO vector containing the gene of interest is int...
- 5pcDNA5/FRT/TO GFP (Plasmid #19444) - Addgene
Plasmid pcDNA5/FRT/TO GFP from Dr. Harm Kampinga's lab contains the insert GFP and is published i...