pTet-On

价格:1200元

联系方式:I47-825O-882O

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pTeton载体基本信息

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: Neomycin_1.html' target='_blank'>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

pTeton载体质粒图谱和多克隆位点信息

pTet-On载体图谱



pTet-On载体特征

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.biofeng.com/
COMMENT     VNTAUTHORNAME|biofeng.com|
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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