pTet-On

pTet-On? expresses the reverse tet-responsive transcriptional activator (rtTA) from the strong immediate early promoter of cytomegalovirus (PCMV). tTA is a fusion of amino acids 1–207 of the tet repressor (TetR) and the negatively charged C-terminal activation domain (130 amino acids) of the VP16 protein of herpes simplex virus. pTet-On is similar to pTet-Off except for four amino acid changes that convert the TetR to a rTetR, and consequently, tTA to rtTA. (pTet-Off also contains several silent mutations.) pTet-On was originally described as pUHD17-1neo by Gossen et al. (1995). pTet-On can be distinguished from pTet-Off by digestion with Hind III.

载体应用

The pTet-On Vector is used to develop stable Tet-On cell lines. After a vector that contains a gene under the control of a tet-responsive element (TRE) is transfected into a Tet-On cell line, the rtTA binds to the TRE, thus activating transcription in the presence of doxycycline (Dox). As Dox is removed from the culture medium, transcription from the TRE is turned off in a highly dose-dependent manner. Additional information on TRE-containing vectors and protocols describing the construction of Tet-On cell lines can be found in the Tet Systems User Manual (PT3001-1).

0001 CTCGAGGAGC TTGGCCCATT GCATACGTTG TATCCATATC ATAATATGTA CATTTATATT 
0061 GGCTCATGTC CAACATTACC GCCATGTTGA CATTGATTAT TGACTAGTTA TTAATAGTAA 
0121 TCAATTACGG GGTCATTAGT TCATAGCCCA TATATGGAGT TCCGCGTTAC ATAACTTACG 
0181 GTAAATGGCC CGCCTGGCTG ACCGCCCAAC GACCCCCGCC CATTGACGTC AATAATGACG 
0241 TATGTTCCCA TAGTAACGCC AATAGGGACT TTCCATTGAC GTCAATGGGT GGAGTATTTA 
0301 CGGTAAACTG CCCACTTGGC AGTACATCAA GTGTATCATA TGCCAAGTAC GCCCCCTATT 
0361 GACGTCAATG ACGGTAAATG GCCCGCCTGG CATTATGCCC AGTACATGAC CTTATGGGAC 
0421 TTTCCTACTT GGCAGTACAT CTACGTATTA GTCATCGCTA TTACCATGGT GATGCGGTTT 
0481 TGGCAGTACA TCAATGGGCG TGGATAGCGG TTTGACTCAC GGGGATTTCC AAGTCTCCAC 
0541 CCCATTGACG TCAATGGGAG TTTGTTTTGG CACCAAAATC AACGGGACTT TCCAAAATGT 
0601 CGTAACAACT CCGCCCCATT GACGCAAATG GGCGGTAGGC GTGTACGGTG GGAGGTCTAT 
0661 ATAAGCAGAG CTCGTTTAGT GAACCGTCAG ATCGCCTGGA GACGCCATCC ACGCTGTTTT 
0721 GACCTCCATA GAAGACACCG GGACCGATCC AGCCTCCGCG GCCCCGAATT CATATGTCTA 
0781 GATTAGATAA AAGTAAAGTG ATTAACAGCG CATTAGAGCT GCTTAATGAG GTCGGAATCG 
0841 AAGGTTTAAC AACCCGTAAA CTCGCCCAGA AGCTTGGTGT AGAGCAGCCT ACACTGTATT 
0901 GGCATGTAAA AAATAAGCGG GCTTTGCTCG ACGCCTTAGC CATTGAGATG TTAGATAGGC 
0961 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