pGBT9

pGBT9 generates a hybrid protein that contains the sequences for the GAL4 DNA-binding domain (DNA-BD; a.a. 1–147). For the construction of a hybrid protein, the gene encoding the protein of interest is ligated into the MCS in the correct orientation and in the correct reading frame such that a fusion protein is generated. The fusion protein is expressed in yeast host cells from the constitutive ADH1 promoter; transcription is terminated at the ADH1 transcription termination signal. The hybrid protein is targeted to the yeast nucleus by nuclear localization sequences that are an intrinsic part of the GAL4 DNA-BD (2). pGBT9 is a shuttle vector that replicates autonomously in both E. coli and S. cerevisiae. It carries the bla gene (for ampicillin resistance in E. coli) and the TRP1 nutritional marker that allow yeast auxotrophs carrying pGBT9 to grow on limiting synthetic medium lacking Trp.

0001 GCTTGCATGC AACTTCTTTT CTTTTTTTTT CTTTTCTCTC TCCCCCGTTG TTGTCTCACC 
0061 ATATCCGCAA TGACAAAAAA AATGATGGAA GACACTAAAG GAAAAAATTA ACGACAAAGA 
0121 CAGCACCAAC AGATGTCGTT GTTCCAGAGC TGATGAGGGG TATCTTCGAA CACACGAAAC 
0181 TTTTTCCTTC CTTCATTCAC GCACACTACT CTCTAATGAG CAACGGTATA CGGCCTTCCT 
0241 TCCAGTTACT TGAATTTGAA ATAAAAAAAG TTTGCCGCTT TGCTATCAAG TATAAATAGA 
0301 CCTGCAATTA TTAATCTTTT GTTTCCTCGT CATTGTTCTC GTTCCCTTTC TTCCTTGTTT 
0361 CTTTTTCTGC ACAATATTTC AAGCTATACC AAGCATACAA TCAACTCCAA GCTTGAAGCA 
0421 AGCCTCCTGA AAGATGAAGC TACTGTCTTC TATCGAACAA GCATGCGATA TTTGCCGACT 
0481 TAAAAAGCTC AAGTGCTCCA AAGAAAAACC GAAGTGCGCC AAGTGTCTGA AGAACAACTG 
0541 GGAGTGTCGC TACTCTCCCA AAACCAAAAG GTCTCCGCTG ACTAGGGCAC ATCTGACAGA 
0601 AGTGGAATCA AGGCTAGAAA GACTGGAACA GCTATTTCTA CTGATTTTTC CTCGAGAAGA 
0661 CCTTGACATG ATTTTGAAAA TGGATTCTTT ACAGGATATA AAAGCATTGT TAACAGGATT 
0721 ATTTGTACAA GATAATGTGA ATAAAGATGC CGTCACAGAT AGATTGGCTT CAGTGGAGAC 
0781 TGATATGCCT CTAACATTGA GACAGCATAG AATAAGTGCG ACATCATCAT CGGAAGAGAG 
0841 TAGTAACAAA GGTCAAAGAC AGTTGACTGT ATCGCCGGAA TTCCCGGGGA TCCGTCGACC 
0901 TGCAGCCAAG CTAATTCCGG GCGAATTTCT TATGATTTAT GATTTTTATT ATTAAATAAG 
0961 TTATAAAAAA AATAAGTGTA TACAAATTTT AAAGTGACTC TTAGGTTTTA AAACGAAAAT 
1021 TCTTGTTCTT GAGTAACTCT TTCCTGTAGG TCAGGTTGCT TTCTCAGGTA TAGCATGAGG 
1081 TCGCTCTTAT TGACCACACC TCTACCGGCA TGCCGGCAAG TGCACAAACA ATACTTAAAT 
1141 AAATACTACT CAGTAATAAC CTATTTCTTA GCATTTTTGA CGAAATTTGC TATTTTGTTA 
1201 GAGTCTTTTA CACCATTTGT CTCCACACCT CCGCTTACAT CAACACCAAT AACGCCATTT 
1261 AATCTAAGCG CATCACCAAC ATTTTCTGGC GTCAGTCCAC CAGCTAACAT AAAATGTAAG 
1321 CTTTCGGGGC TCTCTTGCCT TCCAACCCAG TCAGAAATCG AGTTCCAATC CAAAAGTTCA 
1381 CCTGTCCCAC CTGCTTCTGA ATCAAACAAG GGAATAAACG AATGAGGTTT CTGTGAAGCT 
1441 GCACTGAGTA GTATGTTGCA GTCTTTTGGA AATACGAGTC TTTTAATAAC TGGCAAACCG 
1501 AGGAACTCTT GGTATTCTTG CCACGACTCA TCTCCATGCA GTTGGACGAT ATCAATGCCG 
1561 TAATCATTGA CCAGAGCCAA AACATCCTCC TTAGGTTGAT TACGAAACAC GCCAACCAAG 
1621 TATTTCGGAG TGCCTGAACT ATTTTTATAT GCTTTTACAA GACTTGAAAT TTTCCTTGCA 
1681 ATAACCGGGT CAATTGTTCT CTTTCTATTG GGCACACATA TAATACCCAG CAAGTCAGCA 
1741 TCGGAATCTA GAGCACATTC TGCGGCCTCT GTGCTCTGCA AGCCGCAAAC TTTCACCAAT 
1801 GGACCAGAAC TACCTGTGAA ATTAATAACA GACATACTCC AAGCTGCCTT TGTGTGCTTA 
1861 ATCACGTATA CTCACGTGCT CAATAGTCAC CAATGCCCTC CCTCTTGGCC CTCTCCTTTT 
1921 CTTTTTTCGA CCGAATTAAT TCGTAATCAT GGTCATAGCT GTTTCCTGTG TGAAATTGTT 
1981 ATCCGCTCAC AATTCCACAC AACATACGAG CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG 
2041 CCTAATGAGT GAGGTAACTC ACATTAATTG CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG 
2101 GAAACCTGTC GTGCCAGCTG GATTAATGAA TCGGCCAACG CGCGGGGAGA GGCGGTTTGC 
2161 GTATTGGGCG CTCTTCCGCT TCCTCGCTCA CTGACTCGCT GCGCTCGGTC GTTCGGCTGC 
2221 GGCGAGCGGT ATCAGCTCAC TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA 
2281 ACGCAGGAAA GAACATGTGA GCAAAAGGCC AGCAAAAGGC CAGGAACCGT AAAAAGGCCG 
2341 CGTTGCTGGC GTTTTTCCAT AGGCTCCGCC CCCCTGACGA GCATCACAAA AATCGACGCT 
2401 CAAGTCAGAG GTGGCGAAAC CCGACAGGAC TATAAAGATA CCAGGCGTTT CCCCCTGGAA 
2461 GCTCCCTCGT GCGCTCTCCT GTTCCGACCC TGCCGCTTAC CGGATACCTG TCCGCCTTTC 
2521 TCCCTTCGGG AAGCGTGGCG CTTTCTCATA GCTCACGCTG TAGGTATCTC AGTTCGGTGT 
2581 AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC ACGAACCCCC CGTTCAGCCC GACCGCTGCG 
2641 CCTTATCCGG TAACTATCGT CTTGAGTCCA ACCCGGTAAG ACACGACTTA TCGCCACTGG 
2701 CAGCAGCCAC TGGTAACAGG ATTAGCAGAG CGAGGTATGT AGGCGGTGCT ACAGAGTTCT 
2761 TGAAGTGGTG GCCTAACTAC GGCTACACTA GAAGGACAGT ATTTGGTATC TGCGCTCTGC 
2821 TGAAGCCAGT TACCTTCGGA AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA CAAACCACCG 
2881 CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC 
2941 AAGAAGATCC TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT 
3001 AAGGGATTTT GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA 
3061 AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT 
3121 GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT 
3181 GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG 
3241 CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG 
3301 CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA 
3361 ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG 
3421 CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG 
3481 GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT 
3541 CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA 
3601 TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG 
3661 GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC 
3721 CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG 
3781 GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA 
3841 TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG 
3901 GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT 
3961 GTTGAATACT CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC 
4021 TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA 
4081 CATTTCCCCG AAAAGTGCCA CCTGACGTCT AAGAAACCAT TATTATCATG ACATTAACCT 
4141 ATAAAAATAG GCGTATCACG AGGCCCTTTC GTCTCGCGCG TTTCGGTGAT GACGGTGAAA 
4201 ACCTCTGACA CATGCAGCTC CCGGAGACGG TCACAGCTTG TCTGTAAGCG GATGCCGGGA 
4261 GCAGACAAGC CCGTCAGGGC GCGTCAGCGG GTGTTGGCGG GTGTCGGGGC TGGCTTAACT 
4321 ATGCGGCATC AGAGCAGATT GTACTGAGAG TGCACCATAA CGCATTTAAG CATAAACACG 
4381 CACTATGCCG TTCTTCTCAT GTATATATAT ATACAGGCAA CACGCAGATA TAGGTGCGAC 
4441 GTGAACAGTG AGCTGTATGT GCGCAGCTCG CGTTGCATTT TCGGAAGCGC TCGTTTTCGG 
4501 AAACGCTTTG AAGTTCCTAT TCCGAAGTTC CTATTCTCTA GCTAGAAAGT ATAGGAACTT 
4561 CAGAGCGCTT TTGAAAACCA AAAGCGCTCT GAAGACGCAC TTTCAAAAAA CCAAAAACGC 
4621 ACCGGACTGT AACGAGCTAC TAAAATATTG CGAATACCGC TTCCACAAAC ATTGCTCAAA 
4681 AGTATCTCTT TGCTATATAT CTCTGTGCTA TATCCCTATA TAACCTACCC ATCCACCTTT 
4741 CGCTCCTTGA ACTTGCATCT AAACTCGACC TCTACATTTT TTATGTTTAT CTCTAGTATT 
4801 ACTCTTTAGA CAAAAAAATT GTAGTAAGAA CTATTCATAG AGTGAATCGA AAACAATACG 
4861 AAAATGTAAA CATTTCCTAT ACGTAGTATA TAGAGACAAA ATAGAAGAAA CCGTTCATAA 
4921 TTTTCTGACC AATGAAGAAT CATCAACGCT ATCACTTTCT GTTCACAAAG TATGCGCAAT 
4981 CCACATCGGT ATAGAATATA ATCGGGGATG CCTTTATCTT GAAAAAATGC ACCCGCAGCT 
5041 TCGCTAGTAA TCAGTAAACG CGGGAAGTGG AGTCAGGCTT TTTTTATGGA AGAGAAAATA 
5101 GACACCAAAG TAGCCTTCTT CTAACCTTAA CGGACCTACA GTGCAAAAAG TTATCAAGAG 
5161 ACTGCATTAT AGAGCGCACA AAGGAGAAAA AAAGTAATCT AAGATGCTTT GTTAGAAAAA 
5221 TAGCGCTCTC GGGATGCATT TTTGTAGAAC AAAAAAGAAG TATAGATTCT TTGTTGGTAA 
5281 AATAGCGCTC TCGCGTTGCA TTTCTGTTCT GTAAAAATGC AGCTCAGATT CTTTGTTTGA 
5341 AAAATTAGCG CTCTCGCGTT GCATTTTTGT TTTACAAAAA TGAAGCACAG ATTCTTCGTT 
5401 GGTAAAATAG CGCTTTCGCG TTGCATTTCT GTTCTGTAAA AATGCAGCTC AGATTCTTTG 
5461 TTTGAAAAAT TAGCGCTCTC GCGTTGCATT TTTGTTCTAC AAAATGAAGC ACAGATGCTT 
5521 CGTT