LOCUS       sequence-389375-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-Hook1 fusion protein in 
            yeast (yeast two-hybrid assays) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7712)
  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 7712)
  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-Hook1
FEATURES             Location/Qualifiers
     source          1..7712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        811..1212
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1240..1680
                     /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      3916..4103
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3971..3995)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4156..4830)
                     /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(4831..4932)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4946..4962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4946..4962)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4959..4981)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4970..4986
                     /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(4994..5000,5001..5018,5019..5024))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5039..5060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5177..5194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5348..5936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5428..5447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6107..6898,6899..6967))
                     /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     6730..6749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6968..7072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7140..7158
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7196..7218)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7318..7337
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(7354..7712,1..806)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tataacctac ccatccacct ttcgctcctt gaacttgcat ctaaactcga cctctacatt
       61 ttttatgttt atctctagta ttactcttta gacaaaaaaa ttgtagtaag aactattcat
      121 agagtgaatc gaaaacaata cgaaaatgta aacatttcct atacgtagta tatagagaca
      181 aaatagaaga aaccgttcat aattttctga ccaatgaaga atcatcaacg ctatcacttt
      241 ctgttcacaa agtatgcgca atccacatcg gtatagaata taatcgggga tgcctttatc
      301 ttgaaaaaat gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc
      361 tttttttatg gaagagaaaa tagacaccaa agtagccttc ttctaacctt aacggaccta
      421 cagtgcaaaa agttatcaag agactgcatt atagagcgca caaaggagaa aaaaagtaat
      481 ctaagatgct ttgttagaaa aatagcgctc tcgggatgca tttttgtaga acaaaaaaga
      541 agtatagatt ctttgttggt aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat
      601 gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt gttttacaaa
      661 aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt ctgttctgta
      721 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgttct
      781 acaaaatgaa gcacagatgc ttcgttgctt gcatgcaact tcttttcttt ttttttcttt
      841 tctctctccc ccgttgttgt ctcaccatat ccgcaatgac aaaaaaatga tggaagacac
      901 taaaggaaaa aattaacgac aaagacagca ccaacagatg tcgttgttcc agagctgatg
      961 aggggtatct cgaagcacac gaaacttttt ccttccttca ttcacgcaca ctactctcta
     1021 atgagcaacg gtatacggcc ttccttccag ttacttgaat ttgaaataaa aaaaagtttg
     1081 ctgtcttgct atcaagtata aatagacctg caattattaa tcttttgttt cctcgtcatt
     1141 gttctcgttc cctttcttcc ttgtttcttt ttctgcacaa tatttcaagc tataccaagc
     1201 atacaatcaa ctccaagctt gaagcaagcc tcctgaaaga tgaagctact gtcttctatc
     1261 gaacaagcat gcgatatttg ccgacttaaa aagctcaagt gctccaaaga aaaaccgaag
     1321 tgcgccaagt gtctgaagaa caactgggag tgtcgctact ctcccaaaac caaaaggtct
     1381 ccgctgacta gggcacatct gacagaagtg gaatcaaggc tagaaagact ggaacagcta
     1441 tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga ttctttacag
     1501 gatataaaag cattgttaac aggattattt gtacaagata atgtgaataa agatgccgtc
     1561 acagatagat tggcttcagt ggagactgat atgcctctaa cattgagaca gcatagaata
     1621 agtgcgacat catcatcgga agagagtagt aacaaaggtc aaagacagtt gactgtatcg
     1681 ccggaattcc cggggatcct tatggaggag acgcagccgc cgccgcagcc taagctgccc
     1741 ctgtgcgaca gcctcatgat ctggctgcag acattcaata ctgcctcacc ttgtcaagat
     1801 gtcaaacagc tgactagtgg agttgccatg gcacaagttc ttcatcaaat tgatgcagct
     1861 tggtttaacg aatcttggtt aagccgaatt aaagaggatg ttggggacaa ctggagaata
     1921 aaggccagta atgtaaagaa ggtccttcaa ggaattatga gttattatca tgagtttttg
     1981 gggcagcaga tttcagaagc acttatccct gatttaaacc aaataaccga atgttcagat
     2041 ccagtggagc ttgggaggtt gctccagctt attttaggtt gtgcgatcaa ctgtgaaaag
     2101 aagcaagaac atattcaaaa tataatgaca ctggaagagt ctgttcaaca tgtggtcatg
     2161 actgctattc aagagttgat gagtaaagaa atattgagct ctcctccaaa tgatgctgtt
     2221 ggagaattgg agcaacagct taaaagagcc ttggaagaac ttcaggaagc actagcagaa
     2281 aaagaagagc tgaggcaaag atgtgaagaa ttggatatgc aggtgactac acttcaagat
     2341 gaaaagaatt cactggtttc tgaaaatgag atgatgaatg aaaaacttga ccagttggat
     2401 ggctcttttg atgatccaaa cacagtggtt gcaaaaaagt attttcatgc acaattacaa
     2461 ctagaacaat tacaggaaga aaacttcagg cttgaagctg caaaagatga ttaccgtgtt
     2521 cactgtgaag aacttgaaaa gcagctaatc gaattccagc ataggaatga tgaattgact
     2581 agtcttgcag aagaaacaag agccctgaaa gatgaaatag atgttcttag ggctacctct
     2641 gataaagcaa ataaactgga gtcaacagtt gagatatatc gtcagaagct acaagatctg
     2701 aatgaccttc gcaagcaggt gaaaacttta caggaaacca acatgatgta tatgcataat
     2761 acagtcagct tagaagaaga attaaaaaaa gcaaatgcag cacgtacaca attagaaaca
     2821 tacaaaaggc aggttcaaga tcttcatgtt aaactttcct ccgaatccaa gagggcagac
     2881 acactagcgt ttgaaatgaa gcggcttgaa gaaaaacatg aagctttact taaggaaaaa
     2941 gagagactaa ttgagcagcg tgatactttg aaagaaacaa atgaagagct tcgatgttca
     3001 caagtacaac aggaccacct aaaccaaaca gatgcatctg ctacaaaaag ttatgagaat
     3061 cttgctgctg agattatgcc agtggaatat agggaggtgt ttattcgact gcaacatgaa
     3121 aataagatgc ttcgcttaca gcaagaaggc tctgagaatg aacgtattga ggaacttcag
     3181 gagcagctag aacagaaaca ccgtaaaatg aatgaactgg aaactgagca gaggctgagc
     3241 aaagagcgta ttagagaatt gcagcagcag attgaggacc tccagaaatc tttacaggaa
     3301 caaggttcca agtctgaagg cgaaagttcc agcaaattaa agcagaagtt ggaagctcat
     3361 atggaaaaac tcacagaggt ccatgaagaa ttacagaaga aacaagaact cattgaagat
     3421 cttcagccag atataaatca aaatgtacaa aagatcaatg aacttgaagc tgctcttcag
     3481 aagaaagatg aagatatgaa agcaatggag gaaagatata aaatgtactt ggagaaagcc
     3541 agaaatgtaa taaaaacttt agatcccaag ttaaatccag catcagctga aataatgcta
     3601 ctaagaaagc agttggcaga gaaagagaga agaattgaga ttctggagag tgaatgcaaa
     3661 gtagcaaaat tccgtgatta tgaagaaaaa ctcattgttt ctgcgtggta taataagagt
     3721 ctagcattcc agaaactggg gatggaatct agacttgtga gcggcggtgg tgcctgcagt
     3781 gacactggtg cgtgcactcc tgcgcggtct ttcttagcgc agcaacggca catcaccaac
     3841 accagaagaa atctctctgt taaagtccct gctacaacat ctgattaact cgacctgcag
     3901 ccaagctaat tccgggcgaa tttcttatga tttatgattt ttattattaa ataagttata
     3961 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     4021 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct
     4081 cttattgacc acacctctac cggcatgccg gcaagtgcac aaacaatact taaataaata
     4141 ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc
     4201 ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct
     4261 aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat gtaagctttc
     4321 ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa gttcacctgt
     4381 cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg aagctgcact
     4441 gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca aaccgaggaa
     4501 ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatatcaa tgccgtaatc
     4561 attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa ccaagtattt
     4621 cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc ttgcaataac
     4681 cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt cagcatcgga
     4741 atctagagca cattctgcgg cctctgtgct ctgcaagccg caaactttca ccaatggacc
     4801 agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt gcttaatcac
     4861 gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc cttttctttt
     4921 ttcgaccgaa ttaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     4981 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5041 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5101 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5161 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5221 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5281 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5341 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5401 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5461 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5521 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5581 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5641 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5701 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5761 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5821 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5881 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5941 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6001 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6061 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6241 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7081 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     7141 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     7201 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     7261 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     7321 catcagagca gattgtactg agagtgcacc ataacgcatt taagcataaa cacgcactat
     7381 gccgttcttc tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac
     7441 agtgagctgt atgtgcgcag ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc
     7501 tttgaagttc ctattccgaa gttcctattc tctagctaga aagtatagga acttcagagc
     7561 gcttttgaaa accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa acgcaccgga
     7621 ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct caaaagtatc
     7681 tctttgctat atatctctgt gctatatccc ta
//
