LOCUS       sequence-389850-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-AP-1 gamma-2 ear domain 
            fusion protein in yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  AUTHORS   Mattera R, Arighi CN, Lodge R, Zerial M, Bonifacino JS
  TITLE     Divalent interaction of the GGAs with the Rabaptin-5-Rabex-5 
            complex.
  JOURNAL   EMBO J. 2003 Jan 2;22(1):78-88. doi: 10.1093/emboj/cdg015.
  PUBMED    12505986
REFERENCE   2  (bases 1 to 5867)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "EMBO J"; date: "2003-01-2- 2"; volume: "22"; issue: "1"; pages: 
            "78-88"
COMMENT     Sequence Label: pGBT9-gamma-2 ear
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..865
                     /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"
     terminator      1256..1443
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1311..1335)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1496..2170)
                     /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"
     promoter        complement(2171..2272)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2286..2302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2286..2302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2299..2321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2310..2326
                     /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(join(2334..2340,2341..2358,2359..2364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2379..2400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2517..2534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2688..3276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2768..2787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3447..4238,4239..4307))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4070..4089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4308..4412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4480..4498
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4536..4558)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4658..4677
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4694..5858
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(5863..5867,1..397)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 caacttcttt tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca
       61 atgacaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac
      121 agatgtcgtt gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc
      181 cttcattcac gcacactact ctctaatgag caacggtata cggccttcct tccagttact
      241 tgaatttgaa ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt
      301 attaatcttt tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg
      361 cacaatattt caagctatac caagcataca atcaactcca agcttgaagc aagcctcctg
      421 aaagatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct
      481 caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg
      541 ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc
      601 aaggctagaa agactggaac agctatttct actgattttt cctcgagaag accttgacat
      661 gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca
      721 agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc
      781 tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa
      841 aggtcaaaga cagttgactg tatcgccgga attcctcaaa gtgtttgagc gtgagggagt
      901 acagctgaat ctgtctttca ttcgaccccc tgaaaaccct gctttgctgt taatcaccat
      961 cactgccacc aacttctcag agggtgatgt cacccatttc atctgccagg ctgctgtgcc
     1021 caagagtctc cagctgcagc tgcaggcccc cagtgggaac acagttccag ctcggggtgg
     1081 ccttcctatc acccagctct tcagaatcct caatcctaac aaggcccccc tgcggctaaa
     1141 gctgcgcctc acctacgacc actttcacca gtcggtgcag gagatctttg aggtgaacaa
     1201 cttgcctgtg gaatcgtggc agtaaccgct cgacctgcag ccaagctaat tccgggcgaa
     1261 tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa
     1321 attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct
     1381 gtaggtcagg ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac
     1441 cggcatgccg gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt
     1501 tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca
     1561 cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt
     1621 ctggcgtcag tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa
     1681 cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa
     1741 acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt
     1801 ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg
     1861 actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat
     1921 cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt
     1981 tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc
     2041 tattgggcac acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg
     2101 cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa
     2161 taacagacat actccaagct gcctttgtgt gcttaatcac gtatactcac gtgctcaata
     2221 gtcaccaatg ccctccctct tggccctctc cttttctttt ttcgaccgaa ttaattcgta
     2281 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2401 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2521 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2881 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3121 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3601 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4441 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4501 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4561 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4621 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4681 agagtgcacc ataacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat
     4741 atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag
     4801 ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa
     4861 gttcctattc tctagctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg
     4921 ctctgaagac gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat
     4981 attgcgaata ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt
     5041 gctatatccc tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc
     5101 gacctctaca ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta
     5161 agaactattc atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag
     5221 tatatagaga caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa
     5281 cgctatcact ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg
     5341 gatgccttta tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa
     5401 gtggagtcag gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc
     5461 ttaacggacc tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag
     5521 aaaaaaagta atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta
     5581 gaacaaaaaa gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg
     5641 ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt
     5701 ttgttttaca aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat
     5761 ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg
     5821 catttttgtt ctacaaaatg aagcacagat gcttcgttgc ttgcatg
//
