LOCUS       sequence-389376-        8636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-FHIP-L fusion protein in 
            yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8636)
  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 8636)
  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-FHIP-L
FEATURES             Location/Qualifiers
     source          1..8636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             414..854
                     /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"
     regulatory      1552..1561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      4014..4201
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4069..4093)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4254..4928)
                     /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(4929..5030)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5044..5060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5044..5060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5057..5079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5068..5084
                     /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(5092..5098,5099..5116,5117..5122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5137..5158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5275..5292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5446..6034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5526..5545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6205..6996,6997..7065))
                     /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     6828..6847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7066..7170)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7238..7256
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7294..7316)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7416..7435
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7452..8616
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(8621..8636,1..386)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 cttttttttt cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa
       61 atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg
      121 ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg
      181 cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa
      241 taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt
      301 gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc
      361 aagctatacc aagcatacaa tcaactccaa gcttgaagca agcctcctga aagatgaagc
      421 tactgtcttc tatcgaacaa gcatgcgata tttgccgact taaaaagctc aagtgctcca
      481 aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg ggagtgtcgc tactctccca
      541 aaaccaaaag gtctccgctg actagggcac atctgacaga agtggaatca aggctagaaa
      601 gactggaaca gctatttcta ctgatttttc ctcgagaaga ccttgacatg attttgaaaa
      661 tggattcttt acaggatata aaagcattgt taacaggatt atttgtacaa gataatgtga
      721 ataaagatgc cgtcacagat agattggctt cagtggagac tgatatgcct ctaacattga
      781 gacagcatag aataagtgcg acatcatcat cggaagagag tagtaacaaa ggtcaaagac
      841 agttgactgt atcgccggaa ttcatgatgt catcggtttc gacagaaagc aaactccagc
      901 aggctgtgag cctacaggga gttgacccag aaacatgcat gattgtattt aaaaaccact
      961 gggcacaggt tgtgaaaatc ttggagaagc acgacccctt gaagaacacc caggcaaaat
     1021 atgggtctat ccctccagat gaggccagtg ccgtgcagaa ttacgtagaa cacatgctct
     1081 tcttgttgat tgaagagcaa gccaaagatg ctgcaatggg gccgattctg gaatttgtgg
     1141 tctctgagaa catcatggag aaacttttcc tttggagctt gagaagggag tttactgatg
     1201 agactaaaat tgagcagcta aagatgtatg agatgttggt cacccagtcg caccagcctc
     1261 tgctgcacca caaacccatt ctgaagcctc tgatgatgtt gctgagctct tgttcaggaa
     1321 caaccacccc cactgtggag gagaagctgg ttgtcctact caatcagctc tgttccattc
     1381 ttgccaaaga tccatccatt ttagaactct tcttccacac tagtgaagac caaggcgctg
     1441 ccaacttcct catcttctcc cttctgattc ccttcattca ccgagagggg tcagtaggcc
     1501 agcaagctcg ggatgcattg ctcttcatca tgtctctttc tgctgagaac accatggtgg
     1561 cccatcacat cgtggagaac acctactttt gtccagtact tgcaactggg ctcagtggtc
     1621 tctactcttc cctgcctaca aagctagaag agaaaggcga ggaatggcac tgccttctga
     1681 aagatgactg gcttctactt ccttctcttg tccagttcat gaactccctg gagttttgca
     1741 atgcagtcat acaggtggct caccccttga ttcgaaatca gcttgtcaat tacatttaca
     1801 atggattttt ggtaccagtc ttggctcctg ctctccataa ggtgactgtg gaagaggtca
     1861 tgaccacaac tgcatatctg gaccttttcc tgcgtagcat ctccgagcca gcactacttg
     1921 agatcttcct ccgttttatc ctattgcacc agcacgagaa tgtccacatc ctagacactc
     1981 tcacgagtcg aatcaacacc ccgtttcggc tttgtgtggt gtctctggca ttattcagaa
     2041 ctctcattgg tttacattgt gaagatgtga tgttacagct agttctaagg tatctgatcc
     2101 cctgcaatca catgatgctg agtcagaggt gggctgtgaa ggagagagac tgttactctg
     2161 tttctgcggc caagcttctc gccttgactc ctgtctgctg ctccagcggg atcactctga
     2221 cgctggggaa ccaagagagg gattatattc tctggtcaaa gtgtatgcat gacacttcag
     2281 ggcctgtgga gcggccattc cccgaagcgt tctccgagtc agcctgcatt gtggagtatg
     2341 ggaaagccct ggacatcagc tacctgcagt acctgtggga ggcccacacc aacatcctcc
     2401 gctgcatgag ggactgccgt gtctggtccg ccctgtatga tggcgactcc cccgaccctg
     2461 agatgtttct ccagagtctg acggaggagg gcagtgtgag ctcggcctgc cctgtgttcg
     2521 ggctcccgca acaactcccc aggaagacag gacctcagct ggctcccaga aaggacaaga
     2581 gccagacaga gctggaatgg gatgacagct atgacactgg aatctcctca ggggctgacg
     2641 tgggctcccc agggccttat gatgatctgg aggtttcagg ccccccagca cccattgatc
     2701 cccccaaaca catccaggag atgaagaaga atgccctcct gctcttcaaa gggtcctaca
     2761 tagaagagtc ggactttcag gatgatgtga tggtgtacag gctgtgtgct gagaaggact
     2821 ccgaggacat gaaggattct caggaggaag ctgctaggcc accagctgaa gcccaggctg
     2881 aagttcagag tgtccccatc aacaacggcc ccctcctcag cacccagcca gagacagatt
     2941 cagaggagga gtggaatagg gacaattcag acccgtttca cagtgagccc aaggagccaa
     3001 agcaagagag ggaacctgaa gcagccccag aatccaactc agagttagca tcccctgccc
     3061 ctgaggcaga gcacagctct aacctgacag ccgcccaccc ggagagcgag gagctcattg
     3121 cccagtatga ccaaatcatt aaagagctgg attccggcgc cgagggcttg atggaacaga
     3181 attaccccac acctgatccc ttgcttctca ctaaggagga agaagggaag gaagagagta
     3241 aaggagaaaa ggagaaggag gggaagaagg agctagaaga tgaggaggat gactttgact
     3301 cttttatagc ggagatgcct gctgtagaga ctgtgccttc cccatttgtg gggagagatg
     3361 aggctgcctt tgccagtcgc catcccgtga ggactcaaag caccccattc acaggcccat
     3421 tcatcagcgt agtcctgtca aagctggaga acatgctgga gaactcttta catgttaatt
     3481 tgctgcttat cgggatcatt actcagctag ccagctaccc ccagccactc ctgcgctcct
     3541 ttctgctcaa caccaacatg gtcttccagc caagcgtccg ctctctctat caggtccttg
     3601 catctgtgaa aaacaagatt gaacagtttg cttctgtgga gagagacttc ccagggctcc
     3661 tcattcaagc tcagcagtac ctgctcttcc gtgtggacat gtctgatatg acccctgcag
     3721 cactaaccaa agatcccatt caggaggctt ccaggacagg aagtggcaag aaccttttgg
     3781 atggacctcc aagagtgctt cagcccttcc tgacccacag aaccaaggtg gctgaggcac
     3841 cccccaacct gcccctgccg gtgaggaacc ccatgctggc tgctgccctc ttcccagagt
     3901 tcctgaagga gctggcggcc ttggcccagg aacactccat tctgtgctac aagatcttgg
     3961 gtgactttga ggactcctgc tgttagctcg acctgcagcc aagctaattc cgggcgaatt
     4021 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     4081 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     4141 aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg
     4201 gcatgccggc aagtgcacaa acaatactta aataaatact actcagtaat aacctatttc
     4261 ttagcatttt tgacgaaatt tgctattttg ttagagtctt ttacaccatt tgtctccaca
     4321 cctccgctta catcaacacc aataacgcca tttaatctaa gcgcatcacc aacattttct
     4381 ggcgtcagtc caccagctaa cataaaatgt aagctttcgg ggctctcttg ccttccaacc
     4441 cagtcagaaa tcgagttcca atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac
     4501 aagggaataa acgaatgagg tttctgtgaa gctgcactga gtagtatgtt gcagtctttt
     4561 ggaaatacga gtcttttaat aactggcaaa ccgaggaact cttggtattc ttgccacgac
     4621 tcatctccat gcagttggac gatatcaatg ccgtaatcat tgaccagagc caaaacatcc
     4681 tccttaggtt gattacgaaa cacgccaacc aagtatttcg gagtgcctga actattttta
     4741 tatgctttta caagacttga aattttcctt gcaataaccg ggtcaattgt tctctttcta
     4801 ttgggcacac atataatacc cagcaagtca gcatcggaat ctagagcaca ttctgcggcc
     4861 tctgtgctct gcaagccgca aactttcacc aatggaccag aactacctgt gaaattaata
     4921 acagacatac tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt
     4981 caccaatgcc ctccctcttg gccctctcct tttctttttt cgaccgaatt aattcgtaat
     5041 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5101 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5161 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5221 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5281 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5341 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5401 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5461 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5521 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5581 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5641 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5701 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5761 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5821 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5881 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5941 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6001 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6061 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6121 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6181 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6241 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6301 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6361 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6421 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6481 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6541 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6601 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6661 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6721 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6781 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6841 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6901 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6961 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7021 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7081 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7141 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7201 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7261 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     7321 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     7381 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     7441 agtgcaccat aacgcattta agcataaaca cgcactatgc cgttcttctc atgtatatat
     7501 atatacaggc aacacgcaga tataggtgcg acgtgaacag tgagctgtat gtgcgcagct
     7561 cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt tgaagttcct attccgaagt
     7621 tcctattctc tagctagaaa gtataggaac ttcagagcgc ttttgaaaac caaaagcgct
     7681 ctgaagacgc actttcaaaa aaccaaaaac gcaccggact gtaacgagct actaaaatat
     7741 tgcgaatacc gcttccacaa acattgctca aaagtatctc tttgctatat atctctgtgc
     7801 tatatcccta tataacctac ccatccacct ttcgctcctt gaacttgcat ctaaactcga
     7861 cctctacatt ttttatgttt atctctagta ttactcttta gacaaaaaaa ttgtagtaag
     7921 aactattcat agagtgaatc gaaaacaata cgaaaatgta aacatttcct atacgtagta
     7981 tatagagaca aaatagaaga aaccgttcat aattttctga ccaatgaaga atcatcaacg
     8041 ctatcacttt ctgttcacaa agtatgcgca atccacatcg gtatagaata taatcgggga
     8101 tgcctttatc ttgaaaaaat gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt
     8161 ggagtcaggc tttttttatg gaagagaaaa tagacaccaa agtagccttc ttctaacctt
     8221 aacggaccta cagtgcaaaa agttatcaag agactgcatt atagagcgca caaaggagaa
     8281 aaaaagtaat ctaagatgct ttgttagaaa aatagcgctc tcgggatgca tttttgtaga
     8341 acaaaaaaga agtatagatt ctttgttggt aaaatagcgc tctcgcgttg catttctgtt
     8401 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     8461 gttttacaaa aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt
     8521 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca
     8581 tttttgttct acaaaatgaa gcacagatgc ttcgttgctt gcatgcaact tctttt
//
