出品公司: | Clontech |
---|---|
载体名称: | pTet-On |
质粒类型: | 哺乳细胞表达;四环素调控载体 |
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: | 7.4kb |
5' 测序引物及序列: | -- |
3' 测序引物及序列: | -- |
载体标签: | -- |
载体抗性: | 氨苄青霉素 |
筛选标记: | 新霉素/G418 |
备注: | -- |
产品目录号: | 631018 |
稳定性: | 稳定表达 |
组成型: | 组成型 |
病毒/非病毒: | 非病毒 |
pTet-On哺乳细胞表达;四环素调控载体质粒图谱序列抗性价格报价Biovector NTCC Inc.现货
- 价 格:¥2920
- 货 号:BiovectorpTet-On
- 产 地:北京
- BioVector NTCC典型培养物保藏中心
- 联系人:Dr.Xu, Biovector NTCC Inc.
电话:400-800-2947 工作QQ:1843439339 (微信同号)
邮件:Biovector@163.com
手机:18901268599
地址:北京
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pTeton载体基本信息
pTeton载体质粒图谱和多克隆位点信息
pTeton载体简介
载体描述:
pTet-On表达反向抑制转录激活因子(reverse tet-responsive transcriptional activator, rtTA),该因子来自于巨细胞病毒立即早期启动子(immediate early promoter of cytomegalovirus, Pcmv).tTA是一段融合的1-207个氨基酸序列的Tet抑制子(TetR)和来自于单纯疱疹病毒(Herpes simplex virus,HSV)的VP16蛋白带负电荷C端激活域。
pTet-On与pTet-Off载体质粒相似,除了4个氨基酸的不同。这4个氨基酸的差异使得TetR转变为rTetR,从而使得tTA转变为rtTA. pTet-Off还包含有几个沉默突变。
pTet-on载体被Gossen等(1995)原本描述为pUHD17-1neo.可以通过HindIII限制性内切酶酶切,来区分pTet-On和pTet-Off载体。
载体使用:
pTet-On载体被用来构建稳表达Tet-On细胞系。在将基因插入到载体的Tet抑制子元件(Tet-responsive element, TRE)控制下后,质粒可以用于转染Tet-On细胞系,rtTA可以结合TRE,从而在多西环素(DOX)的存在条件下激活基因转录。DOX从培养基中去除后,TRE调控下的基因转录能够在剂量依赖的条件下下调或者关闭。更多的关于TRE原件包含载体及Tet-On细胞系的操作构建步骤可以参见Tet 系统使用手册(PT3001-1)。
pTeton载体序列
LOCUS pTet-On 7400 bp DNA circular 9-DEC-2009 COMMENT Created by Clontech Laboratories COMMENT Cat. No. 631018 COMMENT This file is created by Vector NTI http://www.biovector.net/ COMMENT VNTAUTHORNAME|biovector.net| FEATURES Location/Qualifiers promoter 86..673 /label=CMV\IE\Promoter CDS 774..1781 /label=rtTA rep_origin 2614..3257 /label=pUC\origin CDS complement(3405..4265) /label=Amp(R) polyA_site 1797..2264 /label=SV40\polyA\signal CDS complement(5677..6471) /label=Neo(R) promoter complement(6791..7134) /label=SV40\early\Promoter BASE COUNT 1876 a 1875 c 1837 g 1812 t ORIGIN 1 ctcgaggagc ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt 61 ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa 121 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 181 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 241 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 301 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 361 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 421 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 481 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 541 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 601 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 661 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt 721 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccccgaatt catatgtcta 781 gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg 841 aaggtttaac aacccgtaaa ctcgcccaga agcttggtgt agagcagcct acactgtatt 901 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc 961 accatactca cttttgccct ttaaaagggg aaagctggca agatttttta cgcaataacg 1021 ctaaaagttt tagatgtgct ttactaagtc atcgcaatgg agcaaaagta cattcagata 1081 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac 1141 aaggtttttc actagagaac gcgttatatg cactcagcgc tgtggggcat tttactttag 1201 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta 1261 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag 1321 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat 1381 gtgaaagtgg gtccgcgtac agccgcgcgc gtacgaaaaa caattacggg tctaccatcg 1441 agggcctgct cgatctcccg gacgacgacg cccccgaaga ggcggggctg gcggctccgc 1501 gcctgtcctt tctccccgcg ggacacacgc gcagactgtc gacggccccc ccgaccgatg 1561 tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg catgccgacg 1621 cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt ccgggattta 1681 ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag tttgagcaga 1741 tgtttaccga tgcccttgga attgacgagt acggtgggta gggggcgcga ggatccagac 1801 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc 1861 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa 1921 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag 1981 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctgcaagc 2041 ctcgtcgtcc tggccggacc acgctatctg tgcaaggtcc ccggccccgg acgcgcgctc 2101 catgagcaga gcgcccgccg ccgaggcgaa gactcgggcg gcgccctgcc cgtcccacca 2161 ggtcaacagg cggtaaccgg cctcttcatc gggaatgcgc gcgaccttca gcatcgccgg 2221 catgtccccc tggcggacgg gaagtatcca gctcgaccaa gcttggcgag attttcagga 2281 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa 2341 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag 2401 accgttcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 2461 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 2521 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 2581 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 2641 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 2701 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 2761 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 2821 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 2881 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 2941 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 3001 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 3061 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 3121 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 3181 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 3241 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 3301 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 3361 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 3421 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 3481 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 3541 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 3601 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 3661 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 3721 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 3781 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 3841 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 3901 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 3961 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 4021 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 4081 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 4141 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 4201 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 4261 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 4321 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 4381 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 4441 aggcgtatca cgaggccctt tcgtcttcac tcgaggtcga gggatccaga catgataaga 4501 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt 4561 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac 4621 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa 4681 agcaagtaaa acctctacaa atgtggtatg gctgattatg atctctagtc aaggcactat 4741 acatcaaata ttccttatta acccctttac aaattaaaaa gctaaaggta cacaattttt 4801 gagcatagtt attaatagca gacactctat gcctgtgtgg agtaagaaaa aacagtatgt 4861 tatgattata actgttatgc ctacttataa aggttacaga atatttttcc ataattttct 4921 tgtatagcag tgcagctttt tcctttgtgg tgtaaatagc aaagcaagca agagttctat 4981 tactaaacac agcatgactc aaaaaactta gcaattctga aggaaagtcc ttggggtctt 5041 ctacctttct cttctttttt ggaggagtag aatgttgaga gtcagcagta gcctcatcat 5101 cactagatgg catttcttct gagcaaaaca ggttttcctc attaaaggca ttccaccact 5161 gctcccattc atcagttcca taggttggaa tctaaaatac acaaacaatt agaatcagta 5221 gtttaacaca ttatacactt aaaaatttta tatttacctt agagctttaa atctctgtag 5281 gtagtttgtc caattatgtc acaccacaga agtaaggttc cttcacaaag atccggacca 5341 aagcggccat cgtgcctccc cactcctgca gttcgggggc atggatgcgc ggatagccgc 5401 tgctggtttc ctggatgccg acggatttgc actgccggta gaactccgcg aggtcgtcca 5461 gcctcaggca gcagctgaac caactcgcga ggggatcgag cccggggtgg gcgaagaact 5521 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga 5581 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg 5641 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg 5701 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc 5761 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata 5821 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac 5881 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat 5941 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag 6001 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt 6061 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc 6121 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg 6181 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc 6241 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt 6301 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg 6361 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct 6421 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga 6481 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa 6541 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg 6601 caattccggt tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc 6661 actgcaagct acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta 6721 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt 6781 cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt gaagctagct ttttgcaaaa 6841 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc 6901 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact 6961 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt 7021 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt 7081 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt 7141 tccacaccct aactgacaca cattccacag ctgcctcgcg cgtttcggtg atgacggtga 7201 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 7261 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat 7321 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag 7381 attgtactga gagtgcacct //
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