LOCUS       sequence-389381-        7667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-Hook2 fusion protein in 
            yeast (yeast two-hybrid assays) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  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 7667)
  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-Hook2
FEATURES             Location/Qualifiers
     source          1..7667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..35)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(189..777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(269..288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(948..1739,1740..1808))
                     /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     1571..1590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1809..1913)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1981..1999
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2037..2059)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2159..2178
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      2195..3359
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3364..3765
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3793..4233
                     /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      6424..6611
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6479..6503)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(6664..7338)
                     /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(7339..7440)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7454..7470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7454..7470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7467..7489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7478..7494
                     /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(7502..7508,7509..7526,7527..7532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7547..7568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
       61 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      121 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      601 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      841 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1081 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1921 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     1981 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     2041 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     2101 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     2161 gcatcagagc agattgtact gagagtgcac cataacgcat ttaagcataa acacgcacta
     2221 tgccgttctt ctcatgtata tatatataca ggcaacacgc agatataggt gcgacgtgaa
     2281 cagtgagctg tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg
     2341 ctttgaagtt cctattccga agttcctatt ctctagctag aaagtatagg aacttcagag
     2401 cgcttttgaa aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg
     2461 actgtaacga gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat
     2521 ctctttgcta tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc
     2581 cttgaacttg catctaaact cgacctctac attttttatg tttatctcta gtattactct
     2641 ttagacaaaa aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat
     2701 gtaaacattt cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc
     2761 tgaccaatga agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca
     2821 tcggtataga atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct
     2881 agtaatcagt aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac
     2941 caaagtagcc ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc
     3001 attatagagc gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg
     3061 ctctcgggat gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag
     3121 cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat
     3181 tagcgctctc gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa
     3241 aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     3301 aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgttg
     3361 cttgcatgca acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca
     3421 tatccgcaat gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca
     3481 gcaccaacag atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt
     3541 tttccttcct tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc
     3601 cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac
     3661 ctgcaattat taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc
     3721 tttttctgca caatatttca agctatacca agcatacaat caactccaag cttgaagcaa
     3781 gcctcctgaa agatgaagct actgtcttct atcgaacaag catgcgatat ttgccgactt
     3841 aaaaagctca agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg
     3901 gagtgtcgct actctcccaa aaccaaaagg tctccgctga ctagggcaca tctgacagaa
     3961 gtggaatcaa ggctagaaag actggaacag ctatttctac tgatttttcc tcgagaagac
     4021 cttgacatga ttttgaaaat ggattcttta caggatataa aagcattgtt aacaggatta
     4081 tttgtacaag ataatgtgaa taaagatgcc gtcacagata gattggcttc agtggagact
     4141 gatatgcctc taacattgag acagcataga ataagtgcga catcatcatc ggaagagagt
     4201 agtaacaaag gtcaaagaca gttgactgta tcgccggaat tcatgagcgt ggacaaagct
     4261 gagctatgcg ggtctctgct cacctggtta cagacgttcc acgttccgtc tccctgtgcc
     4321 agccctcagg acctgagcag cggccttgcc gtagcctatg tgctgaacca gatagacccc
     4381 tcctggttca acgaggcatg gctccagggc atctcggaag atccaggtcc caactggaag
     4441 ctgaaggtca gcaatctgaa gatggtctta cggagcctag tagagtactc ccaggatgtc
     4501 ctggcgcatc ctgtgtcaga agagcatctc ccagatgtga gcctcattgg agagttctca
     4561 gacccggcag agctcggcaa gctgcttcag ctggtgctgg gctgtgccat cagttgcgag
     4621 aaaaagcagg accacatcca gagaatcatg acgctggaag aatcggttca gcatgtggtg
     4681 atggaagcca tccaagagct catgaccaaa gacactcctg actccctgtc accagagacg
     4741 tatggcaact ttgacagcca gtcccgcagg tactatttcc taagtgagga ggctgaggag
     4801 ggggacgaat tacagcagcg ctgtctggat ctggagcggc agctgatgct cctgtcagag
     4861 gagaagcaga gcctggcgca agagaatgca gggctgcggg agcggatggg ccggcctgaa
     4921 ggcgagggta ccccaggtct cactgccaag aagctgctgc tgctgcaatc ccagctggag
     4981 cagttgcagg aggagaactt caggctggag agtggcaggg aggatgagcg cctgcgctgt
     5041 gccgagctgg agagggaggt tgcggagctg cagcaccgga accaggcgct gactagcctg
     5101 gcccaggagg cacaggccct gaaggatgag atggatgaac tacggcagtc ttcggagcgt
     5161 gctgggcagc tggaggccac gctgaccagt tgccggcgcc gcttgggcga gctgagggag
     5221 ctgcggcggc aggtgcggca gctggaggaa cgcaacgccg gccacgccga gcgcacgcga
     5281 caactggagg atgagctacg ccgagcgggc tccctgcgcg cccagctgga ggcgcagcgg
     5341 cggcaggtgc aggaactgca gggccagcgg caggaggagg ccatgaaggc cgagaaatgg
     5401 ctatttgaat gccgcaacct ggaggaaaag tatgagtcgg tgacaaagga gaaggagcgg
     5461 ctgttggcgg agcgggactc cttgcgggag gccaatgagg agctgcgctg cgcccagctg
     5521 cagccgcggg ggttgaccca ggccgatccc tcactggatc ccacctccac acccgtggat
     5581 aacttagccg cagagatcct gcctgcggag ctcagggaga cgctcctgcg gcttcagctg
     5641 gagaacaagc ggctgtgcag gcaggaggcg gccgaccggg agcggcagga ggagctgcag
     5701 cgccagctgg aggatgccaa ccgcgcgcgc cacgggttgg agacgcagca ccggctgaac
     5761 cagcagcagc tatccgagct gcgggcccag gtggaggacc tgcagaaagc cctgcaggag
     5821 caggggggca agactgaaga ttccattttg ctgaaaagga agctggagga acatttgcag
     5881 aagcttcatg aggcagatct ggagttgcag aggaagcggg agtacattga ggagctggag
     5941 ccacccactg acagcagcac agcccggcgg atcgaggagc tgcagcataa cttgcagaag
     6001 aaggacgcgg acttgcgggc catggaggag cgataccgcc gctacgtgga caaggcccgc
     6061 atggtcatgc agaccatgga acccaagcag cggccagctg cgggggcacc tccagaactc
     6121 cattccctga ggacacagct ccgagaacgg gatgtccgca tccgacacct ggagatggac
     6181 tttgagaaaa gccgaagtca gcgggagcag gaagaaaagc tgctcatcag tgcctggtat
     6241 aatatgggca tggccttgca gcagcgagct ggggaggagc gggcgcctgc ccatgcccag
     6301 tcattcctgg cacagcagcg gctggcaacc aattctcgcc gtggaccctt gggacgcctg
     6361 gcatctctga accttcgccc cactgacaag cactgactcg acctgcagcc aagctaattc
     6421 cgggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     6481 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     6541 tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac
     6601 acctctaccg gcatgccggc aagtgcacaa acaatactta aataaatact actcagtaat
     6661 aacctatttc ttagcatttt tgacgaaatt tgctattttg ttagagtctt ttacaccatt
     6721 tgtctccaca cctccgctta catcaacacc aataacgcca tttaatctaa gcgcatcacc
     6781 aacattttct ggcgtcagtc caccagctaa cataaaatgt aagctttcgg ggctctcttg
     6841 ccttccaacc cagtcagaaa tcgagttcca atccaaaagt tcacctgtcc cacctgcttc
     6901 tgaatcaaac aagggaataa acgaatgagg tttctgtgaa gctgcactga gtagtatgtt
     6961 gcagtctttt ggaaatacga gtcttttaat aactggcaaa ccgaggaact cttggtattc
     7021 ttgccacgac tcatctccat gcagttggac gatatcaatg ccgtaatcat tgaccagagc
     7081 caaaacatcc tccttaggtt gattacgaaa cacgccaacc aagtatttcg gagtgcctga
     7141 actattttta tatgctttta caagacttga aattttcctt gcaataaccg ggtcaattgt
     7201 tctctttcta ttgggcacac atataatacc cagcaagtca gcatcggaat ctagagcaca
     7261 ttctgcggcc tctgtgctct gcaagccgca aactttcacc aatggaccag aactacctgt
     7321 gaaattaata acagacatac tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt
     7381 gctcaatagt caccaatgcc ctccctcttg gccctctcct tttctttttt cgaccgaatt
     7441 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7501 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7561 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7621 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtatt
//
