pTRE2

pTRE2 is a response plasmid that can be used to express a gene of interest (Gene X) in Clontech's Tet-OnTM, Tet-OffTM, RevTet-OnTM, and RevTet-OffTM Gene Expression Systems and Cell Lines (1). The Tet Expression Systems and Cell Lines give researchers ready access to the tetracycline-regulated expression systems described by Gossen & Bujard (2; Tet-Off) and Gossen et al. (3; Tet-On). pTRE2 contains an MCS immediately downstream of the Tet-responsive PhCMV*-1 promoter. cDNAs or genes inserted into the MCS will be responsive to the tTA and rtTA regulatory proteins in the Tet-Off and Tet-On systems, respectively. PhCMV*-1 contains the Tet response element (TRE), which consists of seven copies of the 42-bp tet operator sequence (tetO). The TRE is just upstream of the minimal CMV promoter (Pmin CMV), which lacks the enhancer that is part of the complete CMV promoter. Consequently, PhCMV*-1 is silent in the absence of binding of TetR or rTetR to the tetO sequences. Note that the cloned insert must have an initiating ATG codon. In some cases, addition of a Kozak consensus ribosome binding site (4) may improve expression levels; however, many cDNAs have been efficiently expressed in Tet systems without the addition of a Kozak sequence. pTRE2-Gene X plasmids should be cotransfected with the pTK-Hyg Vector (not included) to permit selection of stable transfectants.pTRE2 was originally described as pUHD10-3 in reference 5.

The pTRE2-Luc Control Vector, packaged with the pTRE2 Vector, contains an additional 1649 bp encoding firefly luciferase inserted into the MCS. This vector can be used as a reporter of induction efficiency using standard luciferase detection reagents. It is not intended as a cloning vector.

0001 CTCGAGTTTA CCACTCCCTA TCAGTGATAG AGAAAAGTGA AAGTCGAGTT TACCACTCCC 
0061 TATCAGTGAT AGAGAAAAGT GAAAGTCGAG TTTACCACTC CCTATCAGTG ATAGAGAAAA 
0121 GTGAAAGTCG AGTTTACCAC TCCCTATCAG TGATAGAGAA AAGTGAAAGT CGAGTTTACC 
0181 ACTCCCTATC AGTGATAGAG AAAAGTGAAA GTCGAGTTTA CCACTCCCTA TCAGTGATAG 
0241 AGAAAAGTGA AAGTCGAGTT TACCACTCCC TATCAGTGAT AGAGAAAAGT GAAAGTCGAG 
0301 CTCGGTACCC GGGTCGAGTA GGCGTGTACG GTGGGAGGCC TATATAAGCA GAGCTCGTTT 
0361 AGTGAACCGT CAGATCGCCT GGAGACGCCA TCCACGCTGT TTTGACCTCC ATAGAAGACA 
0421 CCGGGACCGA TCCAGCCTCC GCGGCCCCGA ATTCGAGCTC GGTACCCGGG GATCCTCTAG 
0481 TCAGCTGACG CGTGCTAGCG CGGCCGCATC GATAAGCTTG TCGACGATAT CTCTAGAGCT 
0541 GAGAACTTCA GGGTGAGTTT GGGGACCCTT GATTGTTCTT TCTTTTTCGC TATTGTAAAA 
0601 TTCATGTTAT ATGGAGGGGG CAAAGTTTTC AGGGTGTTGT TTAGAATGGG AAGATGTCCC 
0661 TTGTATCACC ATGGACCCTC ATGATAATTT TGTTTCTTTC ACTTTCTACT CTGTTGACAA 
0721 CCATTGTCTC CTCTTATTTT CTTTTCATTT TCTGTAACTT TTTTCGTTAA ACTTTAGCTT 
0781 GCATTTGTAA CGAATTTTTA AATTCACTTT CGTTTATTTG TCAGATTGTA AGTACTTTCT 
0841 CTAATCACTT TTTTTTCAAG GCAATCAGGG TAATTATATT GTACTTCAGC ACAGTTTTAG 
0901 AGAACAATTG TTATAATTAA ATGATAAGGT AGAATATTTC TGCATATAAA TTCTGGCTGG 
0961 CGTGGAAATA TTCTTATTGG TAGAAACAAC TACATCCTGG TAATCATCCT GCCTTTCTCT 
1021 TTATGGTTAC AATGATATAC ACTGTTTGAG ATGAGGATAA AATACTCTGA GTCCAAACCG 
1081 GGCCCCTCTG CTAACCATGT TCATGCCTTC TTCTTTTTCC TACAGCTCCT GGGCAACGTG 
1141 CTGGTTGTTG TGCTGTCTCA TCATTTTGGC AAAGAATTCA CTCCTCAGGT GCAGGCTGCC 
1201 TATCAGAAGG TGGTGGCTGG TGTGGCCAAT GCCCTGGCTC ACAAATACCA CTGAGATCTT 
1261 TTTCCCTCTG CCAAAAATTA TGGGGACATC ATGAAGCCCC TTGAGCATCT GACTTCTGGG 
1321 TAATAAAGGA AATTTATTTT CATTGCAATA GTGTGTGGGA ATTTTTTGTG TCTCTCACTC 
1381 GGAAGGACAT ATGGGAGGGC AAATCATTTA AAACATCAGA ATGAGTATTT GGTTTAGAGT 
1441 TTGGCAACAT ATGCCATATG CTGGCTGCCA TGAACAAAGG TGGCTATAAA GAGGTCATCA 
1501 GTATATGAAA CAGCCCCCTG CTGTCCATTC CTTATTCCAT AGAAAAGCCT TGACTTGAGG 
1561 TTAGATTTTT TTTATATTTT GTTTTGTGTT ATTTTTTTCT TTAACATCCC TAAAATTTTC 
1621 CTTACATGTT TTACTAGCCA GATTTTTCCT CCTCTCCTGA CTACTCCCAG TCATAGCTGT 
1681 CCCTCTTCTC TTATGAACTC GACTGCATTA ATGAATCGGC CAACGCGCGG GGAGAGGCGG 
1741 TTTGCGTATT GGGCGCTCTT CCGCTTCCTC GCTCACTGAC TCGCTGCGCT CGGTCGTTCG 
1801 GCTGCGGCGA GCGGTATCAG CTCACTCAAA GGCGGTAATA CGGTTATCCA CAGAATCAGG 
1861 GGATAACGCA GGAAAGAACA TGTGAGCAAA AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA 
1921 GGCCGCGTTG CTGGCGTTTT TCCATAGGCT CCGCCCCCCT GACGAGCATC ACAAAAATCG 
1981 ACGCTCAAGT CAGAGGTGGC GAAACCCGAC AGGACTATAA AGATACCAGG CGTTTCCCCC 
2041 TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC GACCCTGCCG CTTACCGGAT ACCTGTCCGC 
2101 CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC TCAATGCTCA CGCTGTAGGT ATCTCAGTTC 
2161 GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG TGTGCACGAA CCCCCCGTTC AGCCCGACCG 
2221 CTGCGCCTTA TCCGGTAACT ATCGTCTTGA GTCCAACCCG GTAAGACACG ACTTATCGCC 
2281 ACTGGCAGCA GCCACTGGTA ACAGGATTAG CAGAGCGAGG TATGTAGGCG GTGCTACAGA 
2341 GTTCTTGAAG TGGTGGCCTA ACTACGGCTA CACTAGAAGG ACAGTATTTG GTATCTGCGC 
2401 TCTGCTGAAG CCAGTTACCT TCGGAAAAAG AGTTGGTAGC TCTTGATCCG GCAAACAAAC 
2461 CACCGCTGGT AGCGGTGGTT TTTTTGTTTG CAAGCAGCAG ATTACGCGCA GAAAAAAAGG 
2521 ATCTCAAGAA GATCCTTTGA TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC 
2581 ACGTTAAGGG ATTTTGGTCA TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTAAA 
2641 TTAAAAATGA AGTTTTAAAT CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA 
2701 CCAATGCTTA ATCAGTGAGG CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT 
2761 TGCCTGACTC CCCGTCGTGT AGATAACTAC GATACGGGAG GGCTTACCAT CTGGCCCCAG 
2821 TGCTGCAATG ATACCGCGAG ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA 
2881 GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC 
2941 TATTAATTGT TGCCGGGAAG CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT 
3001 TGTTGCCATT GCTACAGGCA TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG 
3061 CTCCGGTTCC CAACGATCAA GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT 
3121 TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT 
3181 GGTTATGGCA GCACTGCATA ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT 
3241 GACTGGTGAG TACTCAACCA AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC 
3301 TTGCCCGGCG TCAATACGGG ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT 
3361 CATTGGAAAA CGTTCTTCGG GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG 
3421 TTCGATGTAA CCCACTCGTG CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT 
3481 TTCTGGGTGA GCAAAAACAG GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG 
3541 GAAATGTTGA ATACTCATAC TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA 
3601 TTGTCTCATG AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC 
3661 GCGCACATTT CCCCGAAAAG TGCCACCTGA CGTCTAAGAA ACCATTATTA TCATGACATT 
3721 AACCTATAAA AATAGGCGTA TCACGAGGCC CTTTCGT