LOCUS       Exported                7303 bp DNA     circular SYN 09-SEP-2012
DEFINITION  Yeast two-hybrid ""bait"" vector for expressing proteins fused to 
            the GAL4 DNA-binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    pGBKT7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7303)
  AUTHORS   Clontech
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7303
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1245..1247
                     /codon_start=1
                     /product="start codon for in vitro 
                     transcription/translation"
                     /label=ATG
                     /translation="M"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1281..1333
                     /label=MCS
                     /note="multiple cloning site"
     terminator      1338..1385
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1412..1599
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     primer_bind     complement(1623..1639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1647..1663
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1671..1701)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(2025..2613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2942..2989
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     CDS             complement(3221..4015)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4016..4120)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4147..5489
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5748..6029
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6030..6704
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(6807..7262)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
      421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
      481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
      541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 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
//
