pTet-Off

pTet-Off expresses the tet-responsive transcriptional activator (tTA) 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-Off was originally described as pUHD15-1neo by Resnitzky et al. (1994) and was created by insertion of a neomycin resistance gene into pUHD15-1 (Gossen & Bujard, 1992). pTet-Off can be distinguished from pTet-On by digestion with Hind III.

载体应用

The pTet-Off Vector is used to develop stable Tet-Off cell lines. After a vector that contains a gene under the control of a tet-responsive element (TRE) is transfected into a Tet-Off cell line, the tTA binds to the TRE, thus activating transcription in the absence of tetracycline (Tc), or its derivative doxycycline (Dox). As Tc or Dox is added to the culture medium, transcription from the TRE is turned off in a highly dose-dependent manner. More information on TRE-containing vectors and protocols describing the construction of Tet-Off 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 CGCTAAACTG 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 AGCTAGGTGT AGAGCAGCCT ACATTGTATT 
0901 GGCATGTAAA AAATAAGCGG GCTTTGCTCG ACGCCTTAGC CATTGAGATG TTAGATAGGC 
0961 ACCATACTCA CTTTTGCCCT TTAGAAGGGG AAAGCTGGCA AGATTTTTTA CGTAATAACG 
1021 CTAAAAGTTT TAGATGTGCT TTACTAAGTC ATCGCGATGG AGCAAAAGTA CATTTAGGTA 
1081 CACGGCCTAC AGAAAAACAG TATGAAACTC TCGAAAATCA ATTAGCCTTT TTATGCCAAC 
1141 AAGGTTTTTC ACTAGAGAAT GCATTATATG 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 CTCGTCGTCT GGCCGGACCA CGCTATCTGT GCAAGGTCCC CGGACGCGCG CTCCATGAGC 
2101 AGAGCGCCCG CCGCCGAGGC AAGACTCGGG CGGCGCCCTG CCCGTCCCAC CAGGTCAACA 
2161 GGCGGTAACC GGCCTCTTCA TCGGGAATGC GCGCGACCTT CAGCATCGCC GGCATGTCCC 
2221 CTGGCGGACG GGAAGTATCA GCTCGACCAA GCTTGGCGAG ATTTTCAGGA GCTAAGGAAG 
2281 CTAAAATGGA GAAAAAAATC ACTGGATATA CCACCGTTGA TATATCCCAA TGGCATCGTA 
2341 AAGAACATTT TGAGGCATTT CAGTCAGTTG CTCAATGTAC CTATAACCAG ACCGTTCAGC 
2401 TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCTCTTCCG 
2461 CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT GCGGCGAGCG GTATCAGCTC 
2521 ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA TAACGCAGGA AAGAACATGT 
2581 GAGCAAAAGG CCAGCAAAAG GCCAGGAACC GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC 
2641 ATAGGCTCCG CCCCCCTGAC GAGCATCACA AAAATCGACG CTCAAGTCAG AGGTGGCGAA 
2701 ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG AAGCTCCCTC GTGCGCTCTC 
2761 CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT TCTCCCTTCG GGAAGCGTGG 
2821 CGCTTTCTCA ATGCTCACGC TGTAGGTATC TCAGTTCGGT GTAGGTCGTT CGCTCCAAGC 
2881 TGGGCTGTGT GCACGAACCC CCCGTTCAGC CCGACCGCTG CGCCTTATCC GGTAACTATC 
2941 GTCTTGAGTC CAACCCGGTA AGACACGACT TATCGCCACT GGCAGCAGCC ACTGGTAACA 
3001 GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT CTTGAAGTGG TGGCCTAACT 
3061 ACGGCTACAC TAGAAGGACA GTATTTGGTA TCTGCGCTCT GCTGAAGCCA GTTACCTTCG 
3121 GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC CGCTGGTAGC GGTGGTTTTT 
3181 TTGTTTGCAA GCAGCAGATT ACGCGCAGAA AAAAAGGATC TCAAGAAGAT CCTTTGATCT 
3241 TTTCTACGGG GTCTGACGCT CAGTGGAACG AAAACTCACG TTAAGGGATT TTGGTCATGA 
3301 GATTATCAAA AAGGATCTTC ACCTAGATCC TTTTAAATTA AAAATGAAGT TTTAAATCAA 
3361 TCTAAAGTAT ATATGAGTAA ACTTGGTCTG ACAGTTACCA ATGCTTAATC AGTGAGGCAC 
3421 CTATCTCAGC GATCTGTCTA TTTCGTTCAT CCATAGTTGC CTGACTCCCC GTCGTGTAGA 
3481 TAACTACGAT ACGGGAGGGC TTACCATCTG GCCCCAGTGC TGCAATGATA CCGCGAGACC 
3541 CACGCTCACC GGCTCCAGAT TTATCAGCAA TAAACCAGCC AGCCGGAAGG GCCGAGCGCA 
3601 GAAGTGGTCC TGCAACTTTA TCCGCCTCCA TCCAGTCTAT TAATTGTTGC CGGGAAGCTA 
3661 GAGTAAGTAG TTCGCCAGTT AATAGTTTGC GCAACGTTGT TGCCATTGCT ACAGGCATCG 
3721 TGGTGTCACG CTCGTCGTTT GGTATGGCTT CATTCAGCTC CGGTTCCCAA CGATCAAGGC 
3781 GAGTTACATG ATCCCCCATG TTGTGCAAAA AAGCGGTTAG CTCCTTCGGT CCTCCGATCG 
3841 TTGTCAGAAG TAAGTTGGCC GCAGTGTTAT CACTCATGGT TATGGCAGCA CTGCATAATT 
3901 CTCTTACTGT CATGCCATCC GTAAGATGCT TTTCTGTGAC TGGTGAGTAC TCAACCAAGT 
3961 CATTCTGAGA ATAGTGTATG CGGCGACCGA GTTGCTCTTG CCCGGCGTCA ATACGGGATA 
4021 ATACCGCGCC ACATAGCAGA ACTTTAAAAG TGCTCATCAT TGGAAAACGT TCTTCGGGGC 
4081 GAAAACTCTC AAGGATCTTA CCGCTGTTGA GATCCAGTTC GATGTAACCC ACTCGTGCAC 
4141 CCAACTGATC TTCAGCATCT TTTACTTTCA CCAGCGTTTC TGGGTGAGCA AAAACAGGAA 
4201 GGCAAAATGC CGCAAAAAAG GGAATAAGGG CGACACGGAA ATGTTGAATA CTCATACTCT 
4261 TCCTTTTTCA ATATTATTGA AGCATTTATC AGGGTTATTG TCTCATGAGC GGATACATAT 
4321 TTGAATGTAT TTAGAAAAAT AAACAAATAG GGGTTCCGCG CACATTTCCC CGAAAAGTGC 
4381 CACCTGACGT CTAAGAAACC ATTATTATCA TGACATTAAC CTATAAAAAT AGGCGTATCA 
4441 CGAGGCCCTT TCGTCTTCAC TCGAGGTCGA GGGATCCAGA CATGATAAGA TACATTGATG 
4501 AGTTTGGACA AACCACAACT AGAATGCAGT GAAAAAAATG CTTTATTTGT GAAATTTGTG 
4561 ATGCTATTGC TTTATTTGTA ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT 
4621 GCATTCATTT TATGTTTCAG GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA AGCAAGTAAA 
4681 ACCTCTACAA ATGTGGTATG GCTGATTATG ATCTCTAGTC AAGGCACTAT ACATCAAATA 
4741 TTCCTTATTA ACCCCTTTAC AAATTAAAAA GCTAAAGGTA CACAATTTTT GAGCATAGTT 
4801 ATTAATAGCA GACACTCTAT GCCTGTGTGG AGTAAGAAAA AACAGTATGT TATGATTATA 
4861 ACTGTTATGC CTACTTATAA AGGTTACAGA ATATTTTTCC ATAATTTTCT TGTATAGCAG 
4921 TGCAGCTTTT TCCTTTGTGG TGTAAATAGC AAAGCAAGCA AGAGTTCTAT TACTAAACAC 
4981 AGCATGACTC AAAAAACTTA GCAATTCTGA AGGAAAGTCC TTGGGGTCTT CTACCTTTCT 
5041 CTTCTTTTTT GGAGGAGTAG AATGTTGAGA GTCAGCAGTA GCCTCATCAT CACTAGATGG 
5101 CATTTCTTCT GAGCAAAACA GGTTTTCCTC ATTAAAGGCA TTCCACCACT GCTCCCATTC 
5161 ATCAGTTCCA TAGGTTGGAA TCTAAAATAC ACAAACAATT AGAATCAGTA GTTTAACACA 
5221 TTATACACTT AAAAATTTTA TATTTACCTT AGAGCTTTAA ATCTCTGTAG GTAGTTTGTC 
5281 CAATTATGTC ACACCACAGA AGTAAGGTTC CTTCACAAAG ATCCGGGACC AAAGCGGCCA 
5341 TCGTGCCTCC CCACTCCTGC AGTTCGGGGG CATGGATGCG CGGATAGCCG CTGCTGGTTT 
5401 CCTGGATGCC GACGGATTTG CACTGCCGGT AGAACTCCGC GAGGTCGTCC AGCCTCAGGC 
5461 AGCAGCTGAA CCAACTCGCG AGGGGATCGA GCCCGGGGTG GGCGAAGAAC TCCAGCATGA 
5521 GATCCCCGCG CTGGAGGATC ATCCAGCCGG CGTCCCGGAA AACGATTCCG AAGCCCAACC 
5581 TTTCATAGAA GGCGGCGGTG GAATCGAAAT CTCGTGATGG CAGGTTGGGC GTCGCTTGGT 
5641 CGGTCATTTC GAACCCCAGA GTCCCGCTCA GAAGAACTCG TCAAGAAGGC GATAGAAGGC 
5701 GATGCGCTGC GAATCGGGAG CGGCGATACC GTAAAGCACG AGGAAGCGGT CAGCCCATTC 
5761 GCCGCCAAGC TCTTCAGCAA TATCACGGGT AGCCAACGCT ATGTCCTGAT AGCGGTCCGC 
5821 CACACCCAGC CGGCCACAGT CGATGAATCC AGAAAAGCGG CCATTTTCCA CCATGATATT 
5881 CGGCAAGCAG GCATCGCCAT GGGTCACGAC GAGATCCTCG CCGTCGGGCA TGCGCGCCTT 
5941 GAGCCTGGCG AACAGTTCGG CTGGCGCGAG CCCCTGATGC TCTTCGTCCA GATCATCCTG 
6001 ATCGACAAGA CCGGCTTCCA TCCGAGTACG TGCTCGCTCG ATGCGATGTT TCGCTTGGTG 
6061 GTCGAATGGG CAGGTAGCCG GATCAAGCGT ATGCAGCCGC CGCATTGCAT CAGCCATGAT 
6121 GGATACTTTC TCGGCAGGAG CAAGGTGAGA TGACAGGAGA TCCTGCCCCG GCACTTCGCC 
6181 CAATAGCAGC CAGTCCCTTC CCGCTTCAGT GACAACGTCG AGCACAGCTG CGCAAGGAAC 
6241 GCCCGTCGTG GCCAGCCACG ATAGCCGCGC TGCCTCGTCC TGCAGTTCAT TCAGGGCACC 
6301 GGACAGGTCG GTCTTGACAA AAAGAACCGG GCGCCCCTGC GCTGACAGCC GGAACACGGC 
6361 GGCATCAGAG CAGCCGATTG TCTGTTGTGC CCAGTCATAG CCGAATAGCC TCTCCACCCA 
6421 AGCGGCCGGA GAACCTGCGT GCAATCCATC TTGTTCAATC ATGCGAAACG ATCCTCATCC 
6481 TGTCTCTTGA TCAGATCTTG ATCCCCTGCG CCATCAGATC CTTGGCGGCA AGAAAGCCAT 
6541 CCAGTTTACT TTGCAGGGCT TCCCAACCTT ACCAGAGGGC GCCCCAGCTG GCAATTCCGG 
6601 TTCGCTTGCT GTCCATAAAA CCGCCCAGTC TAGCTATCGC CATGTAAGCC CACTGCAAGC 
6661 TACCTGCTTT CTCTTTGCGC TTGCGTTTTC CCTTGTCCAG ATAGCCCAGT AGCTGACATT 
6721 CATCCGGGGT CAGCACCGTT TCTGCGGACT GGCTTTCTAC GTGTTCCGCT TCCTTTAGCA 
6781 GCCCTTGCGC CCTGAGTGCT TGCGGCAGCG TGTTGCTAGC TTTTTGCAAA AGCCTAGGCC 
6841 TCCAAAAAAG CCTCCTCACT ACTTCTGGAA TAGCTCAGAG GCCGAGGCGG CCTCGGCCTC 
6901 TGCATAAATA AAAAAAATTA GTCAGCCATG GGGCGGAGAA TGGGCGGAAC TGGGCGGAGT 
6961 TAGGGGCGGG ATGGGCGGAG TTAGGGGCGG GACTATGGTT GCTGACTAAT TGAGATGCAT 
7021 GCTTTGCATA CTTCTGCCTG CTGGGGAGCC TGGGGACTTT CCACACCTGG TTGCTGACTA 
7081 ATTGAGATGC ATGCTTTGCA TACTTCTGCC TGCTGGGGAG CCTGGGGACT TTCCACACCC 
7141 TAACTGACAC ACATTCCACA GCTGCCTCGC GCGTTTCGGT GATGACGGTG AAAACCTCTG 
7201 ACACATGCAG CTCCCGGAGA CGGTCACAGC TTGTCTGTAA GCGGATGCCG GGAGCAGACA 
7261 AGCCCGTCAG GGCGCGTCAG CGGGTGTTGG CGGGTGTCGG GGCGCAGCCA TGACCCAGTC 
7321 ACGTAGCGAT AGCGGAGTGT ATACTGGCTT AACTATGCGG CATCAGAGCA GATTGTACTG 
7381 AGAGTGCACC A