The pGBKT7 vector expresses proteins fused to amino acids 1–147 of the GAL4 DNA binding domain (DNA-BD). In yeast, fusion proteins are expressed at high levels from the constitutive ADH1 promoter (PADH1); transcription is terminated by the T7 and ADH1 transcription termination signals (TT7 & ADH1). pGBKT7 also contains the T7 promoter, a c-Myc epitope tag, and a MCS. pGBKT7 replicates autonomously in both E. coli and S. cerevisiae from the pUC and 2 μ ori, respectively. The vector carries the Kanr for selection in E. coli and the TRP1 nutritional marker for selection in yeast. Yeast strains containing pGBKT7 exhibit a higher transformation efficiency than strains carrying other DNA-BD domain vectors (1).

載體應(yīng)用

pGBKT7 is the DNA-BD Vector included with Clontech's Matchmaker? Systems. The MCS of pGBKT7 contains unique restriction sites in frame with the 3' end of the GAL4 DNA-BD for constructing fusion proteins with a bait protein. The bait protein is also expressed as a fusion to a c-Myc epitope tag. c-Myc tagged proteins can be identified with antibodies raised to this common epitope, eliminating the need to generate specific antibodies to new proteins. The T7 promoter is used for in vitro transcription and translation of the epitope tagged fusion protein. Note that the DNA-BD is not expressed during the in vitro transcription and translation reactions.

The MCS in pGBKT7 is compatible with those in pMyc-CMV and pHA-CMV, Clontech's epitope tagged mammalian expression vector set (Cat. No. 631604). As a result, the target gene can be shuttled into these vectors in order to confirm protein interactions in vivo.

0001 CGGTGCGGGC CTCTTCGCTA TTACGCCAGA TCCTTTTGTT GTTTCCGGGT GTACAATATG 
0061 GACTTCCTCT TTTCTGGCAA CCAAACCCAT ACATCGGGAT TCCTATAATA CCTTCGTTGG 
0121 TCTCCCTAAC ATGTAGGTGG CGGAGGGGAG ATATACAATA GAACAGATAC CAGACAAGAC 
0181 ATAATGGGCT AAACAAGACT ACACCAATTA CACTGCCTCA TTGATGGTGG TACATAACGA 
0241 ACTAATACTG TAGCCCTAGA CTTGATAGCC ATCATCATAT CGAAGTTTCA CTACCCTTTT 
0301 TCCATTTGCC ATCTATTGAA GTAATAATAG GCGCATGCAA CTTCTTTTCT TTTTTTTTCT 
0361 TTTCTCTCTC CCCCGTTGTT GTCTCACCAT ATCCGCAATG ACAAAAAAAT GATGGAAGAC 
0421 ACTAAAGGAA AAAATTAACG ACAAAGACAG CACCAACAGA TGTCGTTGTT CCAGAGCTGA 
0481 TGAGGGGTAT CTCGAAGCAC ACGAAACTTT TTCCTTCCTT CATTCACGCA CACTACTCTC 
0541 TAATGAGCAA CGGTATACGG CCTTCCTTCC AGTTACTTGA ATTTGAAATA AAAAAAAGTT 
0601 TGCTGTCTTG CTATCAAGTA TAAATAGACC TGCAATTATT AATCTTTTGT TTCCTCGTCA 
0661 TTGTTCTCGT TCCCTTTCTT CCTTGTTTCT TTTTCTGCAC AATATTTCAA GCTATACCAA 
0721 GCATACAATC AACTCCAAGC TTGAAGCAAG CCTCCTGAAA GATGAAGCTA CTGTCTTCTA 
0781 TCGAACAAGC ATGCGATATT TGCCGACTTA AAAAGCTCAA GTGCTCCAAA GAAAAACCGA 
0841 AGTGCGCCAA GTGTCTGAAG AACAACTGGG AGTGTCGCTA CTCTCCCAAA ACCAAAAGGT 
0901 CTCCGCTGAC TAGGGCACAT CTGACAGAAG TGGAATCAAG GCTAGAAAGA CTGGAACAGC 
0961 TATTTCTACT GATTTTTCCT CGAGAAGACC TTGACATGAT TTTGAAAATG GATTCTTTAC 
1021 AGGATATAAA AGCATTGTTA ACAGGATTAT TTGTACAAGA TAATGTGAAT AAAGATGCCG 
1081 TCACAGATAG ATTGGCTTCA GTGGAGACTG ATATGCCTCT AACATTGAGA CAGCATAGAA 
1141 TAAGTGCGAC ATCATCATCG GAAGAGAGTA GTAACAAAGG TCAAAGACAG TTGACTGTAT 
1201 CGCCGGAATT TGTAATACGA CTCACTATAG GGCGAGCCGC CATCATGGAG GAGCAGAAGC 
1261 TGATCTCAGA GGAGGACCTG CATATGGCCA TGGAGGCCGA ATTCCCGGGG ATCCGTCGAC 
1321 CTGCAGCGGC CGCATAACTA GCATAACCCC TTGGGGCCTC TAAACGGGTC TTGAGGGGTT 
1381 TTTTGCGCGC TTGCAGCCAA GCTAATTCCG GGCGAATTTC TTATGATTTA TGATTTTTAT 
1441 TATTAAATAA GTTATAAAAA AAATAAGTGT ATACAAATTT TAAAGTGACT CTTAGGTTTT 
1501 AAAACGAAAA TTCTTATTCT TGAGTAACTC TTTCCTGTAG GTCAGGTTGC TTTCTCAGGT 
1561 ATAGCATGAG GTCGCTCTTA TTGACCACAC CTCTACCGGC ATGCAAGCTT GGCGTAATCA 
1621 TGGTCATAGC TGTTTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA CAACATACGA 
1681 GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT CACATTAATT 
1741 GCGTTGCGCT CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT GCATTAATGA 
1801 ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC TTCCTCGCTC 
1861 ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG 
1921 GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC 
1981 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC 
2041 CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA CCCGACAGGA 
2101 CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC TGTTCCGACC 
2161 CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT 
2221 AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG 
2281 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG TCTTGAGTCC 
2341 AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG GATTAGCAGA 
2401 GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA CGGCTACACT 
2461 AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT 
2521 GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG 
2581 CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT TTCTACGGGG 
2641 TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT TGGTCATGAG ATTATCAAAA 
2701 AGGATCTTCA CCTAGATCCT TTTAAATTAA AAATGAAGTT TTAAATCAAT CTAAAGTATA 
2761 TATGAGTAAC CTGAGGCTAT GGCAGGGCCT GCCGCCCCGA CGTTGGCTGC GAGCCCTGGG 
2821 CCTTCACCCG AACTTGGGGG GTGGGGTGGG GAAAAGGAAG AAACGCGGGC GTATTGGCCC 
2881 CAATGGGGTC TCGGTGGGGT ATCGACAGAG TGCCAGCCCT GGGACCGAAC CCCGCGTTTA 
2941 TGAACAAACG ACCCAACACC GTGCGTTTTA TTCTGTCTTT TTATTGCCGT CATAGCGCGG 
3001 GTTCCTTCCG GTATTGTCTC CTTCCGTGTT TCAGTTAGCC TCCCCCTAGG GTGGGCGAAG 
3061 AACTCCAGCA TGAGATCCCC GCGCTGGAGG ATCATCCAGC CGGCGTCCCG GAAAACGATT 
3121 CCGAAGCCCA ACCTTTCATA GAAGGCGGCG GTGGAATCGA AATCTCGTGA TGGCAGGTTG 
3181 GGCGTCGCTT GGTCGGTCAT TTCGAACCCC AGAGTCCCGC TCAGAAGAAC TCGTCAAGAA 
3241 GGCGATAGAA GGCGATGCGC TGCGAATCGG GAGCGGCGAT ACCGTAAAGC ACGAGGAAGC 
3301 GGTCAGCCCA TTCGCCGCCA AGCTCTTCAG CAATATCACG GGTAGCCAAC GCTATGTCCT 
3361 GATAGCGGTC CGCCACACCC AGCCGGCCAC AGTCGATGAA TCCAGAAAAG CGGCCATTTT 
3421 CCACCATGAT ATTCGGCAAG CAGGCATCGC CATGAGTCAC GACGAGATCC TCGCCGTCGG 
3481 GCATGCTCGC CTTGAGCCTG GCGAACAGTT CGGCTGGCGC GAGCCCCTGA TGCTCTTCGT 
3541 CCAGATCATC CTGATCGACA AGACCGGCTT CCATCCGAGT ACGTGCTCGC TCGATGCGAT 
3601 GTTTCGCTTG GTGGTCGAAT GGGCAGGTAG CCGGATCAAG CGTATGCAGC CGCCGCATTG 
3661 CATCAGCCAT GATGGATACT TTCTCGGCAG GAGCAAGGTG AGATGACAGG AGATCCTGCC 
3721 CCGGCACTTC GCCCAATAGC AGCCAGTCCC TTCCCGCTTC AGTGACAACG TCGAGCACAG 
3781 CTGCGCAAGG AACGCCCGTC GTGGCCAGCC ACGATAGCCG CGCTGCCTCG TCTTGCAGTT 
3841 CATTCAGGGC ACCGGACAGG TCGGTCTTGA CAAAAAGAAC CGGGCGCCCC TGCGCTGACA 
3901 GCCGGAACAC GGCGGCATCA GAGCAGCCGA TTGTCTGTTG TGCCCAGTCA TAGCCGAATA 
3961 GCCTCTCCAC CCAAGCGGCC GGAGAACCTG CGTGCAATCC ATCTTGTTCA ATCATACTCT 
4021 TCCTTTTTCA ATATTATTGA AGCATTTATC AGGGTTATTG TCTCATGAGC GGATACATAT 
4081 TTGAATGTAT TTAGAAAAAT AAACAAATAG GGGTTCCGCG CACATTTCCC CGAAAAGTGC 
4141 CACCTGAACG AAGCATCTGT GCTTCATTTT GTAGAACAAA AATGCAACGC GAGAGCGCTA 
4201 ATTTTTCAAA CAAAGAATCT GAGCTGCATT TTTACAGAAC AGAAATGCAA CGCGAAAGCG 
4261 CTATTTTACC AACGAAGAAT CTGTGCTTCA TTTTTGTAAA ACAAAAATGC AACGCGAGAG 
4321 CGCTAATTTT TCAAACAAAG AATCTGAGCT GCATTTTTAC AGAACAGAAA TGCAACGCGA 
4381 GAGCGCTATT TTACCAACAA AGAATCTATA CTTCTTTTTT GTTCTACAAA AATGCATCCC 
4441 GAGAGCGCTA TTTTTCTAAC AAAGCATCTT AGATTACTTT TTTTCTCCTT TGTGCGCTCT 
4501 ATAATGCAGT CTCTTGATAA CTTTTTGCAC TGTAGGTCCG TTAAGGTTAG AAGAAGGCTA 
4561 CTTTGGTGTC TATTTTCTCT TCCATAAAAA AAGCCTGACT CCACTTCCCG CGTTTACTGA 
4621 TTACTAGCGA AGCTGCGGGT GCATTTTTTC AAGATAAAGG CATCCCCGAT TATATTCTAT 
4681 ACCGATGTGG ATTGCGCATA CTTTGTGAAC AGAAAGTGAT AGCGTTGATG ATTCTTCATT 
4741 GGTCAGAAAA TTATGAACGG TTTCTTCTAT TTTGTCTCTA TATACTACGT ATAGGAAATG 
4801 TTTACATTTT CGTATTGTTT TCGATTCACT CTATGAATAG TTCTTACTAC AATTTTTTTG 
4861 TCTAAAGAGT AATACTAGAG ATAAACATAA AAAATGTAGA GGTCGAGTTT AGATGCAAGT 
4921 TCAAGGAGCG AAAGGTGGAT GGGTAGGTTA TATAGGGATA TAGCACAGAG ATATATAGCA 
4981 AAGAGATACT TTTGAGCAAT GTTTGTGGAA GCGGTATTCG CAATATTTTA GTAGCTCGTT 
5041 ACAGTCCGGT GCGTTTTTGG TTTTTTGAAA GTGCGTCTTC AGAGCGCTTT TGGTTTTCAA 
5101 AAGCGCTCTG AAGTTCCTAT ACTTTCTAGA GAATAGGAAC TTCGGAATAG GAACTTCAAA 
5161 GCGTTTCCGA AAACGAGCGC TTCCGAAAAT GCAACGCGAG CTGCGCACAT ACAGCTCACT 
5221 GTTCACGTCG CACCTATATC TGCGTGTTGC CTGTATATAT ATATACATGA GAAGAACGGC 
5281 ATAGTGCGTG TTTATGCTTA AATGCGTACT TATATGCGTC TATTTATGTA GGATGAAAGG 
5341 TAGTCTAGTA CCTCCTGTGA TATTATCCCA TTCCATGCGG GGTATCGTAT GCTTCCTTCA 
5401 GCACTACCCT TTAGCTGTTC TATATGCTGC CACTCCTCAA TTGGATTAGT CTCATCCTTC 
5461 AATGCTATCA TTTCCTTTGA TATTGGATCA TACTAAGAAA CCATTATTAT CATGACATTA 
5521 ACCTATAAAA ATAGGCGTAT CACGAGGCCC TTTCGTCTCG CGCGTTTCGG TGATGACGGT 
5581 GAAAACCTCT GACACATGCA GCTCCCGGAG ACGGTCACAG CTTGTCTGTA AGCGGATGCC 
5641 GGGAGCAGAC AAGCCCGTCA GGGCGCGTCA GCGGGTGTTG GCGGGTGTCG GGGCTGGCTT 
5701 AACTATGCGG CATCAGAGCA GATTGTACTG AGAGTGCACC ATAGATCAAC GACATTACTA 
5761 TATATATAAT ATAGGAAGCA TTTAATAGAA CAGCATCGTA ATATATGTGT ACTTTGCAGT 
5821 TATGACGCCA GATGGCAGTA GTGGAAGATA TTCTTTATTG AAAAATAGCT TGTCACCTTA 
5881 CGTACAATCT TGATCCGGAG CTTTTCTTTT TTTGCCGATT AAGAATTAAT TCGGTCGAAA 
5941 AAAGAAAAGG AGAGGGCCAA GAGGGAGGGC ATTGGTGACT ATTGAGCACG TGAGTATACG 
6001 TGATTAAGCA CACAAAGGCA GCTTGGAGTA TGTCTGTTAT TAATTTCACA GGTAGTTCTG 
6061 GTCCATTGGT GAAAGTTTGC GGCTTGCAGA GCACAGAGGC CGCAGAATGT GCTCTAGATT 
6121 CCGATGCTGA CTTGCTGGGT ATTATATGTG TGCCCAATAG AAAGAGAACA ATTGACCCGG 
6181 TTATTGCAAG GAAAATTTCA AGTCTTGTAA AAGCATATAA AAATAGTTCA GGCACTCCGA 
6241 AATACTTGGT TGGCGTGTTT CGTAATCAAC CTAAGGAGGA TGTTTTGGCT CTGGTCAATG 
6301 ATTACGGCAT TGATATCGTC CAACTGCATG GAGATGAGTC GTGGCAAGAA TACCAAGAGT 
6361 TCCTCGGTTT GCCAGTTATT AAAAGACTCG TATTTCCAAA AGACTGCAAC ATACTACTCA 
6421 GTGCAGCTTC ACAGAAACCT CATTCGTTTA TTCCCTTGTT TGATTCAGAA GCAGGTGGGA 
6481 CAGGTGAACT TTTGGATTGG AACTCGATTT CTGACTGGGT TGGAAGGCAA GAGAGCCCCG 
6541 AAAGCTTACA TTTTATGTTA GCTGGTGGAC TGACGCCAGA AAATGTTGGT GATGCGCTTA 
6601 GATTAAATGG CGTTATTGGT GTTGATGTAA GCGGAGGTGT GGAGACAAAT GGTGTAAAAG 
6661 ACTCTAACAA AATAGCAAAT TTCGTCAAAA ATGCTAAGAA ATAGGTTATT ACTGAGTAGT 
6721 ATTTATTTAA GTATTGTTTG TGCACTTGCC GATCTATGCG GTGTGAAATA CCGCACAGAT 
6781 GCGTAAGGAG AAAATACCGC ATCAGGAAAT TGTAAGCGTT AATATTTTGT TAAAATTCGC 
6841 GTTAAATTTT TGTTAAATCA GCTCATTTTT TAACCAATAG GCCGAAATCG GCAAAATCCC 
6901 TTATAAATCA AAAGAATAGA CCGAGATAGG GTTGAGTGTT GTTCCAGTTT GGAACAAGAG 
6961 TCCACTATTA AAGAACGTGG ACTCCAACGT CAAAGGGCGA AAAACCGTCT ATCAGGGCGA 
7021 TGGCCCACTA CGTGAACCAT CACCCTAATC AAGTTTTTTG GGGTCGAGGT GCCGTAAAGC 
7081 ACTAAATCGG AACCCTAAAG GGAGCCCCCG ATTTAGAGCT TGACGGGGAA AGCCGGCGAA 
7141 CGTGGCGAGA AAGGAAGGGA AGAAAGCGAA AGGAGCGGGC GCTAGGGCGC TGGCAAGTGT 
7201 AGCGGTCACG CTGCGCGTAA CCACCACACC CGCCGCGCTT AATGCGCCGC TACAGGGCGC 
7261 GTCCATTCGC CATTCAGGCT GCGCAACTGT TGGGAAGGGC GAT