LOCUS       sequence-387523-        6461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-AAGAB fusion protein in 
            yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6461)
  AUTHORS   Mattera R, De Pace R, Bonifacino JS
  TITLE     The adaptor protein chaperone AAGAB stabilizes AP-4 complex 
            subunits.
  JOURNAL   Mol Biol Cell. 2022 Oct 1;33(12):ar109. doi: 
            10.1091/mbc.E22-05-0177. Epub 2022 Aug 17.
  PUBMED    35976721
REFERENCE   2  (bases 1 to 6461)
  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"; journalName: "Mol Biol 
            Cell."; date: "2022-10-1"; volume: "33(12):ar109. doi"; pages: " 
            10.1091/mbc.E22-05-0177. Epub 2022 Aug 17"
COMMENT     Sequence Label: pGBT9-AAGAB
FEATURES             Location/Qualifiers
     source          1..6461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             434..874
                     /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      1859..2046
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1914..1938)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2099..2773)
                     /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(2774..2875)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2889..2905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2889..2905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2902..2924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2913..2929
                     /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(2937..2943,2944..2961,2962..2967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2982..3003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3120..3137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3291..3879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3371..3390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4050..4841,4842..4910))
                     /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     4673..4692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4911..5015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5083..5101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5139..5161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5261..5280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      5297..6461
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac
      121 agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact
      181 ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt
      241 ccagttactt gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca
      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
      841 tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcatggctg ctggcgtacc
      901 ctgtgcgtta gtcaccagct gctcctccgt cttctcagga gaccagctgg tccaacatat
      961 ccttggaaca gaagatctta ttgtggaagt gacttccaat gatgctgtga gattttatcc
     1021 ctggaccatt gataataaat actattcagc agacatcaat ctatgtgtgg tgccaaacaa
     1081 atttcttgtt actgcagaga ttgcagaatc tgtccaagca tttgtggttt actttgacag
     1141 cacacaaaaa tcgggccttg atagtgtctc ctcatggctt ccactggcaa aagcatggtt
     1201 acctgaggtg atgatcttgg tctgcgatag agtgtctgaa gatggtataa accgacaaaa
     1261 agctcaagaa tggtgcatca aacatggctt tgaattggta gaacttagtc cagaggagtt
     1321 gcctgaggag gatgatgact tcccagaatc tacaggagta aagcgaattg tccaagccct
     1381 gaatgccaat gtgtggtcca atgtagtgat gaagaatgat aggaaccaag gctttagcct
     1441 tctcaactca ttgactggaa caaaccatag cattgggtca gcagatccct gtcacccaga
     1501 gcaaccccat ttgccagcag cagatagtac tgaatccctc tctgatcatc ggggtggtgc
     1561 atctaacaca acagatgccc aggttgatag cattgtggat cccatgttag atctggatat
     1621 tcaagaatta gccagtctta ccactggagg aggagatgtg gagaattttg aaagactctt
     1681 ttcaaagtta aaggaaatga aagacaaggc tgcgacgctt cctcatgagc aaagaaaagt
     1741 gcatgcagaa aaggtggcca aagcattctg gatggcaatc gggggagaca gagatgaaat
     1801 tgaaggcctt tcatctgatg aagagcactg actcgacctg cagccaagct aattccgggc
     1861 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
     1921 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
     1981 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc
     2041 taccggcatg ccggcaagtg cacaaacaat acttaaataa atactactca gtaataacct
     2101 atttcttagc atttttgacg aaatttgcta ttttgttaga gtcttttaca ccatttgtct
     2161 ccacacctcc gcttacatca acaccaataa cgccatttaa tctaagcgca tcaccaacat
     2221 tttctggcgt cagtccacca gctaacataa aatgtaagct ttcggggctc tcttgccttc
     2281 caacccagtc agaaatcgag ttccaatcca aaagttcacc tgtcccacct gcttctgaat
     2341 caaacaaggg aataaacgaa tgaggtttct gtgaagctgc actgagtagt atgttgcagt
     2401 cttttggaaa tacgagtctt ttaataactg gcaaaccgag gaactcttgg tattcttgcc
     2461 acgactcatc tccatgcagt tggacgatat caatgccgta atcattgacc agagccaaaa
     2521 catcctcctt aggttgatta cgaaacacgc caaccaagta tttcggagtg cctgaactat
     2581 ttttatatgc ttttacaaga cttgaaattt tccttgcaat aaccgggtca attgttctct
     2641 ttctattggg cacacatata atacccagca agtcagcatc ggaatctaga gcacattctg
     2701 cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg accagaacta cctgtgaaat
     2761 taataacaga catactccaa gctgcctttg tgtgcttaat cacgtatact cacgtgctca
     2821 atagtcacca atgccctccc tcttggccct ctccttttct tttttcgacc gaattaattc
     2881 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2941 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3001 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3121 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3181 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3481 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3721 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3961 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4021 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4081 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4141 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4201 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4261 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4321 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4381 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4441 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4501 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4561 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4621 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4681 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4741 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4801 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4861 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4921 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4981 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5041 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5101 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5161 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5221 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5281 ctgagagtgc accataacgc atttaagcat aaacacgcac tatgccgttc ttctcatgta
     5341 tatatatata caggcaacac gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg
     5401 cagctcgcgt tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc
     5461 gaagttccta ttctctagct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa
     5521 gcgctctgaa gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa
     5581 aatattgcga ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc
     5641 tgtgctatat ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa
     5701 ctcgacctct acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta
     5761 gtaagaacta ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg
     5821 tagtatatag agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat
     5881 caacgctatc actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc
     5941 ggggatgcct ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg
     6001 gaagtggagt caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta
     6061 accttaacgg acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag
     6121 gagaaaaaaa gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt
     6181 gtagaacaaa aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt
     6241 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca
     6301 tttttgtttt acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg
     6361 catttctgtt ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg
     6421 ttgcattttt gttctacaaa atgaagcaca gatgcttcgt t
//
