LOCUS       sequence-393106-        8438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-FHIP fusion protein in 
            yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  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 8438)
  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
FEATURES             Location/Qualifiers
     source          1..8438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(102..119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(273..861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(353..372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1032..1823,1824..1892))
                     /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     1655..1674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1893..1997)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2065..2083
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2121..2143)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2243..2262
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      2279..3443
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3448..3849
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3877..4317
                     /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      7279..7466
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(7334..7358)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(7519..8193)
                     /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(8194..8295)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8309..8325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8309..8325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8322..8344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8333..8349
                     /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(8357..8363,8364..8381,8382..8387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8402..8423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
       61 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      241 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
      961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1021 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1081 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1141 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1201 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1261 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1321 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1381 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1441 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1501 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1561 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1621 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1681 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     1741 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     1801 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     1861 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     1921 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     1981 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2041 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     2101 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2161 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2221 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataac
     2281 gcatttaagc ataaacacgc actatgccgt tcttctcatg tatatatata tacaggcaac
     2341 acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc gttgcatttt
     2401 cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc tattctctag
     2461 ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg aagacgcact
     2521 ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc gaataccgct
     2581 tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat atccctatat
     2641 aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct ctacattttt
     2701 tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac tattcataga
     2761 gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat agagacaaaa
     2821 tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta tcactttctg
     2881 ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc ctttatcttg
     2941 aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga gtcaggcttt
     3001 ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac ggacctacag
     3061 tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa aagtaatcta
     3121 agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca aaaaagaagt
     3181 atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg taaaaatgca
     3241 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ttacaaaaat
     3301 gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg ttctgtaaaa
     3361 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttctaca
     3421 aaatgaagca cagatgcttc gttgcttgca tgcaacttct tttctttttt tttcttttct
     3481 ctctcccccg ttgttgtctc accatatccg caatgacaaa aaaatgatgg aagacactaa
     3541 aggaaaaaat taacgacaaa gacagcacca acagatgtcg ttgttccaga gctgatgagg
     3601 ggtatctcga agcacacgaa actttttcct tccttcattc acgcacacta ctctctaatg
     3661 agcaacggta tacggccttc cttccagtta cttgaatttg aaataaaaaa aagtttgctg
     3721 tcttgctatc aagtataaat agacctgcaa ttattaatct tttgtttcct cgtcattgtt
     3781 ctcgttccct ttcttccttg tttctttttc tgcacaatat ttcaagctat accaagcata
     3841 caatcaactc caagcttgaa gcaagcctcc tgaaagatga agctactgtc ttctatcgaa
     3901 caagcatgcg atatttgccg acttaaaaag ctcaagtgct ccaaagaaaa accgaagtgc
     3961 gccaagtgtc tgaagaacaa ctgggagtgt cgctactctc ccaaaaccaa aaggtctccg
     4021 ctgactaggg cacatctgac agaagtggaa tcaaggctag aaagactgga acagctattt
     4081 ctactgattt ttcctcgaga agaccttgac atgattttga aaatggattc tttacaggat
     4141 ataaaagcat tgttaacagg attatttgta caagataatg tgaataaaga tgccgtcaca
     4201 gatagattgg cttcagtgga gactgatatg cctctaacat tgagacagca tagaataagt
     4261 gcgacatcat catcggaaga gagtagtaac aaaggtcaaa gacagttgac tgtatcgccg
     4321 gaattcatgg agaggatgaa ttggctgagc agactggcct cccggggccc tgggcaccgt
     4381 atacctcaag gggccaatct ccaaacccca gtcatggctg atcccgagac ctgcctcatg
     4441 gtcttcaaga atcactggtc ccaggtggtg cgaatcctgg agcggcaagg ccctcgggca
     4501 gctcctgggg gtgcagacga tctcagtgct gtgcgcaacc acacttacca gatgttgaca
     4561 ctgctggcag aggaccgtgc agttccctcg gcccccacag gccctgggcc cctgctggag
     4621 tttgctctgc acgaggatct gctgacccgt gtgttgacat ggcagctgca atgggatgag
     4681 cttggggatg gggtcgagga acggcgggct gagcaactga aactatttga aatgctagtg
     4741 agcgaagctc gccagccact gttgcggcat ggtccagttc gtgaggctct gctcaccctg
     4801 ctggatgcct gtggccgccc tgtgcccagt agcccagcac tggatgaagg cttggtgcta
     4861 cttctcagcc agctgtgtgt ttgtgtggcc caggagcctt cattgctcga gttcttcctg
     4921 cagccacctc ctgagcctgg agccgctccc cgtcttcttc tcttttctcg ccttgtccct
     4981 tttgtgcatc gagagggcac cctgggccag caggcccgtg atgccctact tcttctcatg
     5041 gctttgtcag ctgggagccc cactgtgggc cgctacatcg cggatcactc ttacttctgc
     5101 ccggtgctgg ccacagggct cagtgccctg tactcatcac tgcctcgaaa gattgaggtt
     5161 ccaggggatg attggcactg tctgcgacgg gaagactggc tgggagtgcc agcccttgca
     5221 ctcttcatga gttccctgga gttctgcaat gcagtaattc aggtggctca ccccctggtg
     5281 cagaagcagt tggttgatta tatccataat gggttcctgg tgcctgtcat gggtcctgcc
     5341 ttgcacaaga cctctgtgga ggagatgatc gccagtaccg cctacctgga acttttccta
     5401 cggagtatct cagagcctgc tttgctccgt accttcctgc gattcctgtt gttgcaccgg
     5461 catgacaccc acaccatcct cgacaccctc gttgctcgta ttggcagtaa ctcccggctc
     5521 tgcatggtct ctctgagtct cttcaggacc ctcctgaacc tcagctgtga ggatgtcctg
     5581 ctgcagctgg ttctcaggta tcttgttcca tgtaaccacg ttatgctgag ccagaagccg
     5641 gctgttcgtg atgtggacct atatggacga gcagctgaca agtttctctc cctaatccca
     5701 cgctgttgtc ggcaccacgc ccccagccca cctcgtccag agcatgcctc atgggcacga
     5761 ggtcctggaa gcccaagtgt ggactcctct tctgtgacga cagtaccccg gccctccaca
     5821 ccatctcgtc tggctctctt cctgcggcag cagagcctgg gtggctctga gtctccaggc
     5881 ccagcccctt gctcaccagg gctttctgca tcccccgcct ccagccctgg ccgacggcct
     5941 acccctgcag aggagcctgg agagctggaa gacaattacc tggagtatct gcgtgaggca
     6001 cgtcgtggtg tggaccgctg tgtccgagcc tgccgtacct ggtctgcccc ctatgatggc
     6061 gagcggccct ctcctgagcc cagtcctttt ggctcccgga ctaagaaacg cagcctactg
     6121 catgaggagg acaggaacaa cgtgggggaa ggggaggagg aagagctggg gaggaggggg
     6181 cgggctgggg gtgcagggga gggccctggt cacctgcccc ctccccagct caatggagtg
     6241 ccaggatcat ggcctgaggg ggccaagaag gttcgtttgg tgccaaagga gggagctggg
     6301 gaactgctag agggcatctc cgagggcatg gcaggactag agggctttgg gcaggagctc
     6361 cgggagctag aggtggcatt gagcaatggg ggaactggct cagagtcccc cttagaacct
     6421 ccactgcccc ttgaggagga ggaggcctac gagagcttca cctgtccccc tgagccccct
     6481 ggccccttcc tcagcagccc tttgcggact ctcaaccagc tgccaagcca gcccttcact
     6541 ggccccttca tggctgtgct ctttgccaaa ctcgagaaca tgctgcagaa ctccgtctat
     6601 gtcaacttcc tgctgacggg gctggtggcc cagctggcct gtcaccccca gcccctgctc
     6661 cgctctttcc tgctcaacac caacatggtc ttccagccca gtgtcaagtc cctgctgcag
     6721 gtgctgggct ctgtgaagaa taagattgag aactttgcgg cttcccagga ggacttccca
     6781 gcactgctgt ccaaagccaa gaagtacctc attgcccgtg gcaagttgga ctgggctgag
     6841 ggccctgcag caggacctgc cccacgccgt tctgatcccc tagtgaagag ccggaggcca
     6901 tccttggggg agttactcct gcggcatgca cacagtccaa ccagggcccg gcaggcggca
     6961 caattggtcc ttcagcctgg gcgagacgga gcaggacttg gcctaagtgg gggctcccct
     7021 ggggcttcaa ctccagttct actcacccgg ggcggggccc ctgaacgcca aggtgaggct
     7081 cttcgagtca agaatgctgt ctactgtgca gtcattttcc ctgaatttct caaggagttg
     7141 gctgccatct cccaggctca tgccgtcacc tcgcctttct tgttggagac ttcagaggaa
     7201 ggatctggcc ctctcatctc aggctgtggg cccctcaatc cttaaggatc cgtcgacctg
     7261 cagccaagct aattccgggc gaatttctta tgatttatga tttttattat taaataagtt
     7321 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     7381 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc
     7441 gctcttattg accacacctc taccggcatg ccggcaagtg cacaaacaat acttaaataa
     7501 atactactca gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga
     7561 gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa
     7621 tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagct
     7681 ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca aaagttcacc
     7741 tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct gtgaagctgc
     7801 actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg gcaaaccgag
     7861 gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat caatgccgta
     7921 atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc caaccaagta
     7981 tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt tccttgcaat
     8041 aaccgggtca attgttctct ttctattggg cacacatata atacccagca agtcagcatc
     8101 ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg
     8161 accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg tgtgcttaat
     8221 cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct ctccttttct
     8281 tttttcgacc gaattaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     8341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     8401 aatgagtgag ctaactcaca ttaattgcgt tgcgctca
//
