LOCUS       sequence-389860-        5911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-GGA2-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 5911)
  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 5911)
  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-GGA2-GAE
FEATURES             Location/Qualifiers
     source          1..5911
                     /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      1300..1487
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1355..1379)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1540..2214)
                     /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(2215..2316)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2330..2346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2330..2346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2343..2365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2354..2370
                     /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(2378..2384,2385..2402,2403..2408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2423..2444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2561..2578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2732..3320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2812..2831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3491..4282,4283..4351))
                     /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     4114..4133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4352..4456)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4524..4542
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4580..4602)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4702..4721
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4738..5902
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(5907..5911,1..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 attcccgggg atccccctca ttgtgtatga
      901 ccggaatgga ttcagaattc tgctccactt ctcccagacg ggagcccctg ggcacccaga
      961 ggtacaggtg ctgctcttga ccatgatgag cacggctccc cagcctgtct gggatatcat
     1021 gtttcaagtg gctgtgccaa agtcaatgag agtgcagctg cagccggcat ccagctccaa
     1081 gcttcctgca ttcagtcctt tgatgcctcc agctgtgata tctcagatgc tgctgcttga
     1141 caatccacac aaagaaccta tccgcttacg gtacaagctg acattcaacc aaggtggaca
     1201 gcctttcagc gaagtaggag aagtgaaaga cttcccagac ctggctgtct tgggcgcagc
     1261 ctaagcggcc gcgtcgacct gcagccaagc taattccggg cgaatttctt atgatttatg
     1321 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
     1381 taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt
     1441 tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat gccggcaagt
     1501 gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag catttttgac
     1561 gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc cgcttacatc
     1621 aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg tcagtccacc
     1681 agctaacata aaatgtaagc tttcggggct ctcttgcctt ccaacccagt cagaaatcga
     1741 gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga
     1801 atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa atacgagtct
     1861 tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat ctccatgcag
     1921 ttggacgata tcaatgccgt aatcattgac cagagccaaa acatcctcct taggttgatt
     1981 acgaaacacg ccaaccaagt atttcggagt gcctgaacta tttttatatg cttttacaag
     2041 acttgaaatt ttccttgcaa taaccgggtc aattgttctc tttctattgg gcacacatat
     2101 aatacccagc aagtcagcat cggaatctag agcacattct gcggcctctg tgctctgcaa
     2161 gccgcaaact ttcaccaatg gaccagaact acctgtgaaa ttaataacag acatactcca
     2221 agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc aatgccctcc
     2281 ctcttggccc tctccttttc ttttttcgac cgaattaatt cgtaatcatg tcatagctgt
     2341 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2461 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2521 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4501 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     4561 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4621 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4681 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataacg
     4741 catttaagca taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca
     4801 cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc
     4861 ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctagc
     4921 tagaaagtat aggaacttca gagcgctttt gaaaaccaaa agcgctctga agacgcactt
     4981 tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta aaatattgcg aataccgctt
     5041 ccacaaacat tgctcaaaag tatctctttg ctatatatct ctgtgctata tccctatata
     5101 acctacccat ccacctttcg ctccttgaac ttgcatctaa actcgacctc tacatttttt
     5161 atgtttatct ctagtattac tctttagaca aaaaaattgt agtaagaact attcatagag
     5221 tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac gtagtatata gagacaaaat
     5281 agaagaaacc gttcataatt ttctgaccaa tgaagaatca tcaacgctat cactttctgt
     5341 tcacaaagta tgcgcaatcc acatcggtat agaatataat cggggatgcc tttatcttga
     5401 aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag tcaggctttt
     5461 tttatggaag agaaaataga caccaaagta gccttcttct aaccttaacg gacctacagt
     5521 gcaaaaagtt atcaagagac tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa
     5581 gatgctttgt tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta
     5641 tagattcttt gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag
     5701 ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt tacaaaaatg
     5761 aagcacagat tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa
     5821 tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa
     5881 aatgaagcac agatgcttcg ttgcttgcat g
//
