LOCUS       sequence-389379-        6392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-FTS fusion protein in 
            yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6392)
  AUTHORS   Mattera R, Williamson CD, Ren X, Bonifacino JS
  TITLE     The FTS-Hook-FHIP (FHF) complex interacts with AP-4 to mediate 
            perinuclear distribution of AP-4 and its cargo ATG9A.
  JOURNAL   Mol Biol Cell. 2020 Apr 15;31(9):963-979. doi: 
            10.1091/mbc.E19-11-0658. Epub 2020 Feb 19.
  PUBMED    32073997
REFERENCE   2  (bases 1 to 6392)
  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: 
            "10.1091/mbc.E19-11-0658"; journalName: "Mol Biol Cell"; date: 
            "2020-04-15- 15"; volume: "31"; issue: "9"; pages: "963-979"
COMMENT     Sequence Label: pGBT9-FTS
FEATURES             Location/Qualifiers
     source          1..6392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..440
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             468..908
                     /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      1824..2011
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1879..1903)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2064..2738)
                     /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(2739..2840)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2854..2870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2854..2870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2867..2889)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2878..2894
                     /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(2902..2908,2909..2926,2927..2932))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2947..2968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3085..3102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3256..3844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3336..3355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4015..4806,4807..4875))
                     /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     4638..4657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4876..4980)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5048..5066
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5104..5126)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5226..5245
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(5262..6392,1..34)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tttgttctac aaaatgaagc acagatgctt cgttgcttgc atgcaacttc ttttcttttt
       61 ttttcttttc tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaatgatg
      121 gaagacacta aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag
      181 agctgatgag gggtatctcg aagcacacga aactttttcc ttccttcatt cacgcacact
      241 actctctaat gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa
      301 aaagtttgct gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc
      361 tcgtcattgt tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta
      421 taccaagcat acaatcaact ccaagcttga agcaagcctc ctgaaagatg aagctactgt
      481 cttctatcga acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc tccaaagaaa
      541 aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg tcgctactct cccaaaacca
      601 aaaggtctcc gctgactagg gcacatctga cagaagtgga atcaaggcta gaaagactgg
      661 aacagctatt tctactgatt tttcctcgag aagaccttga catgattttg aaaatggatt
      721 ctttacagga tataaaagca ttgttaacag gattatttgt acaagataat gtgaataaag
      781 atgccgtcac agatagattg gcttcagtgg agactgatat gcctctaaca ttgagacagc
      841 atagaataag tgcgacatca tcatcggaag agagtagtaa caaaggtcaa agacagttga
      901 ctgtatcgcc ggaattcatg aaccctttct ggagcatgtc tacaagctct gtacgcaaac
      961 gatctgaagg tgaagagaag acattaacag gggacgtgaa aaccagtcct ccacgaactg
     1021 caccaaagaa acagctgcct tctattccca aaaatgcttt gcccataact aagcctacat
     1081 ctcctgcccc agcagcacag tcaacaaatg gcacgcatgc gtcctatgga cccttctacc
     1141 tggaatactc tcttcttgca gaatttacct tggttgtgaa gcagaagcta ccaggcgtct
     1201 atgtgcagcc atcttatcgc tctgcattaa tgtggtttgg agtaatattc atacggcatg
     1261 gactttacca agatggcgta tttaagttta cagtttacat ccctgataac tatccagatg
     1321 gtgactgtcc acgcttggtg ttcgatattc ctgtctttca cccgctagtt gatcccacct
     1381 caggtgagct ggatgtgaag agagcatttg caaaatggag gcggaaccat aatcatattt
     1441 ggcaggtatt aatgtatgca aggagagttt tctacaagat tgatacagca agccccctga
     1501 acccagaggc tgcagtactg tatgaaaaag atattcagct ttttaaaagt aaagttgttg
     1561 acagtgttaa ggtgtgcact gctcgtttgt ttgaccaacc taaaatagaa gacccctatg
     1621 caattagctt ttctccatgg aatccttctg tacatgatga agccagagaa aagatgctga
     1681 ctcagaaaaa gcctgaagaa cagcacaata aaagtgttca tgttgctggc ctgtcatggg
     1741 taaagcctgg ctcagtacag cctttcagta aagaagagaa aacagtggcg acttaactcg
     1801 acctgcagcc aagctaattc cgggcgaatt tcttatgatt tatgattttt attattaaat
     1861 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     1921 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg
     1981 aggtcgctct tattgaccac acctctaccg gcatgccggc aagtgcacaa acaatactta
     2041 aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg
     2101 ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca
     2161 tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt
     2221 aagctttcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca atccaaaagt
     2281 tcacctgtcc cacctgcttc tgaatcaaac aagggaataa acgaatgagg tttctgtgaa
     2341 gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat aactggcaaa
     2401 ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac gatatcaatg
     2461 ccgtaatcat tgaccagagc caaaacatcc tccttaggtt gattacgaaa cacgccaacc
     2521 aagtatttcg gagtgcctga actattttta tatgctttta caagacttga aattttcctt
     2581 gcaataaccg ggtcaattgt tctctttcta ttgggcacac atataatacc cagcaagtca
     2641 gcatcggaat ctagagcaca ttctgcggcc tctgtgctct gcaagccgca aactttcacc
     2701 aatggaccag aactacctgt gaaattaata acagacatac tccaagctgc ctttgtgtgc
     2761 ttaatcacgt atactcacgt gctcaatagt caccaatgcc ctccctcttg gccctctcct
     2821 tttctttttt cgaccgaatt aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     2881 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2941 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3061 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4981 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5041 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5101 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5161 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5221 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aacgcattta agcataaaca
     5281 cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga tataggtgcg
     5341 acgtgaacag tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc
     5401 ggaaacgctt tgaagttcct attccgaagt tcctattctc tagctagaaa gtataggaac
     5461 ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
     5521 gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
     5581 aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
     5641 ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt atctctagta
     5701 ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc gaaaacaata
     5761 cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga aaccgttcat
     5821 aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa agtatgcgca
     5881 atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat gcacccgcag
     5941 cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa
     6001 tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa agttatcaag
     6061 agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa
     6121 aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt ctttgttggt
     6181 aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga ttctttgttt
     6241 gaaaaattag cgctctcgcg ttgcattttt gttttacaaa aatgaagcac agattcttcg
     6301 ttggtaaaat agcgctttcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     6361 tgtttgaaaa attagcgctc tcgcgttgca tt
//
