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pTet-On载体 pTeton质粒基本信息
pTet-On质粒图谱 pTeton载体图谱和pTet-On载体序列 pTeton质粒序列多克隆位点信息
pTet-On质粒 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）。
pTet-On质粒序列 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
//
