LOCUS       sequence-389831-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-GGA1-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 5891)
  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 5891)
  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-GGA1-GAE
FEATURES             Location/Qualifiers
     source          1..5891
                     /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      1280..1467
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1335..1359)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1520..2194)
                     /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(2195..2296)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2310..2326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2310..2326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2323..2345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2334..2350
                     /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(2358..2364,2365..2382,2383..2388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2403..2424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2541..2558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2712..3300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2792..2811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3471..4262,4263..4331))
                     /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     4094..4113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4332..4436)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4504..4522
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4560..4582)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4682..4701
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4718..5882
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(5887..5891,1..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 attcgtgact gtgtatgacc agcacggctt
      901 ccgcatcctc ttccattttg cccgggaccc actgccaggg cgctccgacg tgctggtggt
      961 ggtggtttcc atgctgagca ccgcccccca gcccatccgc aacatcgtgt tccagtcagc
     1021 tgtccccaag gttatgaagg tgaagctgca gccaccctcg ggcacggagc tgccagcttt
     1081 taaccccatc gtccacccct cagcaatcac ccaggtcctg ctgcttgcca acccccagaa
     1141 ggagaaggtt cgcctccgct acaagctcac cttcaccatg ggtgaccaga cctacaacga
     1201 gatgggggat gtggaccagt tccccccacc tgaaacctgg ggtagcctct gagtcgacct
     1261 gcagccaagc taattccggg cgaatttctt atgatttatg atttttatta ttaaataagt
     1321 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt
     1381 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt
     1441 cgctcttatt gaccacacct ctaccggcat gccggcaagt gcacaaacaa tacttaaata
     1501 aatactactc agtaataacc tatttcttag catttttgac gaaatttgct attttgttag
     1561 agtcttttac accatttgtc tccacacctc cgcttacatc aacaccaata acgccattta
     1621 atctaagcgc atcaccaaca ttttctggcg tcagtccacc agctaacata aaatgtaagc
     1681 tttcggggct ctcttgcctt ccaacccagt cagaaatcga gttccaatcc aaaagttcac
     1741 ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga atgaggtttc tgtgaagctg
     1801 cactgagtag tatgttgcag tcttttggaa atacgagtct tttaataact ggcaaaccga
     1861 ggaactcttg gtattcttgc cacgactcat ctccatgcag ttggacgata tcaatgccgt
     1921 aatcattgac cagagccaaa acatcctcct taggttgatt acgaaacacg ccaaccaagt
     1981 atttcggagt gcctgaacta tttttatatg cttttacaag acttgaaatt ttccttgcaa
     2041 taaccgggtc aattgttctc tttctattgg gcacacatat aatacccagc aagtcagcat
     2101 cggaatctag agcacattct gcggcctctg tgctctgcaa gccgcaaact ttcaccaatg
     2161 gaccagaact acctgtgaaa ttaataacag acatactcca agctgccttt gtgtgcttaa
     2221 tcacgtatac tcacgtgctc aatagtcacc aatgccctcc ctcttggccc tctccttttc
     2281 ttttttcgac cgaattaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     2341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2521 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4441 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4501 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4561 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4621 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4681 gcggcatcag agcagattgt actgagagtg caccataacg catttaagca taaacacgca
     4741 ctatgccgtt cttctcatgt atatatatat acaggcaaca cgcagatata ggtgcgacgt
     4801 gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc gttttcggaa
     4861 acgctttgaa gttcctattc cgaagttcct attctctagc tagaaagtat aggaacttca
     4921 gagcgctttt gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc aaaaacgcac
     4981 cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat tgctcaaaag
     5041 tatctctttg ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg
     5101 ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac
     5161 tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa
     5221 aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt
     5281 ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc
     5341 acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc
     5401 gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga
     5461 caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac
     5521 tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata
     5581 gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta tagattcttt gttggtaaaa
     5641 tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa
     5701 aattagcgct ctcgcgttgc atttttgttt tacaaaaatg aagcacagat tcttcgttgg
     5761 taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt
     5821 tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa aatgaagcac agatgcttcg
     5881 ttgcttgcat g
//
