LOCUS       sequence-389387-        7670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-Hook3 fusion protein in 
            yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7670)
  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 7670)
  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-Hook3
FEATURES             Location/Qualifiers
     source          1..7670
                     /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      3874..4061
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3929..3953)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4114..4788)
                     /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(4789..4890)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4904..4920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4904..4920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4917..4939)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4928..4944
                     /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(4952..4958,4959..4976,4977..4982))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4997..5018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5135..5152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5306..5894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5386..5405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6065..6856,6857..6925))
                     /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     6688..6707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6926..7030)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7098..7116
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7154..7176)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7276..7295
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(7312..7670,1..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 ccggaattca tgttcagcgt agagtcgctg gagcgggcgg agctgtgcga gagcctcctc
     1741 acttggatcc agacatttaa tgtggatgca ccatgccaga ccgtggaaga tttaacgaat
     1801 ggggttgtga tggcccaggt tcttcaaaag atagatcctg catattttga tgaaaattgg
     1861 ctaaacagaa tcaaaactga agtaggagat aattggaggc taaagataag caatttaaag
     1921 aaaattttaa aaggaatctt ggattataat catgagattt taggacagca aattaatgac
     1981 tttacccttc ctgatgtgaa ccttattggg gagcattctg atgcagcaga gcttggaagg
     2041 atgcttcagc tcatcttagg ctgtgctgtg aactgtgaac agaagcaaga gtacatccaa
     2101 gccattatga tgatggagga atctgttcaa catgttgtca tgacagccat tcaagagctg
     2161 atgagtaaag aatctcctgt ctctgctgga aatgatgcct atgttgacct tgatcgtcag
     2221 ctgaagaaaa ctacagagga actaaatgaa gctttgtcag caaaggaaga aattgctcaa
     2281 agatgccatg aactggatat gcaggttgca gcattgcagg aagagaaaag tagtttgttg
     2341 gcagagaatc aggtattaat ggaaagactc aatcaatctg attctataga agaccctaac
     2401 agtccagcag gaagaaggca tttgcagctc cagactcaat tagaacagct ccaagaagaa
     2461 acattcagac tagaagcagc caaagatgat tatcgaatac gttgtgaaga gttagaaaag
     2521 gagatctctg aacttcggca acagaatgat gaactgacca ctttggcaga tgaagctcag
     2581 tctctgaaag atgagatcga cgtgctgaga cattcttctg ataaagtatc taaactagaa
     2641 ggtcaagtag aatcttataa aaagaagcta gaagaccttg gtgatttaag gcggcaggtt
     2701 aaactcttag aagagaagaa taccatgtat atgcagaata ctgtcagtct agaggaagag
     2761 ttaagaaagg ccaacgcagc gcgaagtcaa cttgaaacct acaagagaca ggtagtagaa
     2821 ctacaaaaca gattatccga agaatcaaag aaagcagata aactagattt tgaatataag
     2881 cggctaaaag aaaaagttga cagtcttcaa aaagaaaagg acaggctgag aacagaaagg
     2941 gattctctga aggaaaccat tgaagagctt cgttgtgtac aagctcaaga agggcagctc
     3001 acaacacaag ggttaatgcc tcttggaagt caggagtctt cagacagtct agctgcagag
     3061 attgttacac ctgaaatcag ggagaaactt attcgtcttc agcatgagaa taagatgtta
     3121 aagcttaacc aagaaggttc ggacaatgaa aaaatagcct tattgcagag ccttctagat
     3181 gatgcaaatc tacgcaagaa tgaactggag acagagaata ggctggtgaa tcaaagactt
     3241 ctggaagtac agtcacaagt tgaagaatta caaaaatctt tacaggatca aggctcaaaa
     3301 gcagaagatt cagtccttct aaaaaagaag cttgaagaac atctagagaa gctgcatgag
     3361 gccaataatg aactacagaa gaagagagcc attattgaag atctcgagcc aagatttaac
     3421 aacagctcct taaaaattga agaattacaa gaagctttac gaaagaaaga ggaagaaatg
     3481 aagcaaatgg aagaacgata caaaaaatac ttagagaaag ccaaaagtgt catccgtact
     3541 ttagatccta aacagaatca aggagcagca ccagaaatac aagctcttaa aaatcagctc
     3601 caggaacgag accgactgtt ccactcatta gagaaagaat atgagaaaac aaagagtcag
     3661 agagagatgg aagagaaata tattgttagt gcctggtaca atatgggaat gaccctgcat
     3721 aaaaaggcag ctgaagatag actggcaagc acaggctcag ggcagtcatt tctggcgagg
     3781 cagaggcaag cgaccagcag cagaagatca tacccaggcc acgtgcagcc ggccacagca
     3841 aggtaggtcg acctgcagcc aagctaattc cgggcgaatt tcttatgatt tatgattttt
     3901 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     3961 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     4021 gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgccggc aagtgcacaa
     4081 acaatactta aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt
     4141 tgctattttg ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc
     4201 aataacgcca tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa
     4261 cataaaatgt aagctttcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca
     4321 atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac aagggaataa acgaatgagg
     4381 tttctgtgaa gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat
     4441 aactggcaaa ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac
     4501 gatatcaatg ccgtaatcat tgaccagagc caaaacatcc tccttaggtt gattacgaaa
     4561 cacgccaacc aagtatttcg gagtgcctga actattttta tatgctttta caagacttga
     4621 aattttcctt gcaataaccg ggtcaattgt tctctttcta ttgggcacac atataatacc
     4681 cagcaagtca gcatcggaat ctagagcaca ttctgcggcc tctgtgctct gcaagccgca
     4741 aactttcacc aatggaccag aactacctgt gaaattaata acagacatac tccaagctgc
     4801 ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt caccaatgcc ctccctcttg
     4861 gccctctcct tttctttttt cgaccgaatt aattcgtaat catgtcatag ctgtttcctg
     4921 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4981 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5041 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5101 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5161 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5221 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5281 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5341 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5401 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5461 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5521 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5581 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5641 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5701 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5761 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5821 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5881 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5941 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6001 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6061 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6121 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6181 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6241 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6301 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6361 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6421 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6481 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6541 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6601 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6661 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6721 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6781 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6841 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6901 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6961 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7021 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7081 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     7141 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7201 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7261 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aacgcattta
     7321 agcataaaca cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga
     7381 tataggtgcg acgtgaacag tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc
     7441 gctcgttttc ggaaacgctt tgaagttcct attccgaagt tcctattctc tagctagaaa
     7501 gtataggaac ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa
     7561 aaccaaaaac gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa
     7621 acattgctca aaagtatctc tttgctatat atctctgtgc tatatcccta
//
