LOCUS       sequence-389855-        6212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-GGA3-GAE 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 6212)
  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 6212)
  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-GGA3-GAE
FEATURES             Location/Qualifiers
     source          1..6212
                     /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      1601..1788
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1656..1680)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1841..2515)
                     /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(2516..2617)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2631..2647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2631..2647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2644..2666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2655..2671
                     /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(2679..2685,2686..2703,2704..2709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2724..2745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2862..2879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3033..3621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3113..3132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3792..4583,4584..4652))
                     /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     4415..4434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4653..4757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4825..4843
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4881..4903)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5003..5022
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      5039..6203
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(6208..6212,1..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 attcccaaaa gttgagcccg cagtccctgg
      901 gcaccatggc ttggcgttgg gcaacagcgc gctgcaccac ctggatgccc tcgatcagct
      961 tctagaagag gccaaagtga cctcgggctt ggtgaaaccc actacctccc ctctcatccc
     1021 caccaccacc ccagccaggc ccctcctgcc cttctccacg gggcccggca gcccgctctt
     1081 ccagccactg agtttccagt cccagggcag ccccccgaag gggcctgagc tctccctggc
     1141 cagcatccac gtgcccctgg aatcgatcaa gcctagcagt gcccttcctg tgacagccta
     1201 cgataaaaac ggcttccgca tcctcttcca ctttgccaag gagtgtcccc caggacgacc
     1261 tgacgtgctg gtggtggtgg tgtccatgct gaacacggct cccttacctg tcaagagcat
     1321 cgtgctgcag gctgcagtgc ccaagtcaat gaaagtgaag ttgcagccac cttctgggac
     1381 agaactctct ccatttagcc ccatccagcc acctgcagcc atcacccagg tcatgttgct
     1441 ggccaatcca ctgaaggaga aggtgcggct tcggtataag ctgaccttcg ccctggggga
     1501 gcagctgagc acagaggtgg gcgaggtgga ccagttccct cctgtggaac agtgggggaa
     1561 cctatgaaga tccgtcgacc tgcagccaag ctaattccgg gcgaatttct tatgatttat
     1621 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     1681 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     1741 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccggcaag
     1801 tgcacaaaca atacttaaat aaatactact cagtaataac ctatttctta gcatttttga
     1861 cgaaatttgc tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat
     1921 caacaccaat aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac
     1981 cagctaacat aaaatgtaag ctttcggggc tctcttgcct tccaacccag tcagaaatcg
     2041 agttccaatc caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg
     2101 aatgaggttt ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc
     2161 ttttaataac tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca
     2221 gttggacgat atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat
     2281 tacgaaacac gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa
     2341 gacttgaaat tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata
     2401 taatacccag caagtcagca tcggaatcta gagcacattc tgcggcctct gtgctctgca
     2461 agccgcaaac tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc
     2521 aagctgcctt tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc
     2581 cctcttggcc ctctcctttt cttttttcga ccgaattaat tcgtaatcat gtcatagctg
     2641 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2701 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2761 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2821 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4741 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4801 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4861 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4921 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4981 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataac
     5041 gcatttaagc ataaacacgc actatgccgt tcttctcatg tatatatata tacaggcaac
     5101 acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc gttgcatttt
     5161 cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc tattctctag
     5221 ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg aagacgcact
     5281 ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc gaataccgct
     5341 tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat atccctatat
     5401 aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct ctacattttt
     5461 tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac tattcataga
     5521 gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat agagacaaaa
     5581 tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta tcactttctg
     5641 ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc ctttatcttg
     5701 aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga gtcaggcttt
     5761 ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac ggacctacag
     5821 tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa aagtaatcta
     5881 agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca aaaaagaagt
     5941 atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg taaaaatgca
     6001 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ttacaaaaat
     6061 gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg ttctgtaaaa
     6121 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttctaca
     6181 aaatgaagca cagatgcttc gttgcttgca tg
//
