LOCUS       sequence-387538-        8695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 transcriptional activation domain (AD)-AP-3 
            sigma-3B fusion protein in yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8695)
  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 8695)
  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: pACT2-AP-3 sigma-3B
FEATURES             Location/Qualifiers
     source          1..8695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..438
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             484..504
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             520..861
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             874..900
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             880..906
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     terminator      1912..2099
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1967..1991)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(2120..2153)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2231..2264)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2308..2341)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2419..2452)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2961..4055)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4056..4461)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      4938..6280
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5901..5948)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        6545..6649
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6650..6718,6719..7510)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6868..6887)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7681..8269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8170..8189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8423..8440
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8455..8595
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8521..8540)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(8681..8695,1..8)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcaacttctt ttcttttttt
       61 ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa aaatgatgga
      121 agacactaaa ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt tgttccagag
      181 ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt ccttcattca cgcacactac
      241 tctctaatga gcaacggtat acggccttcc ttccagttac ttgaatttga aataaaaaaa
      301 agtttgctgt cttgctatca agtataaata gacctgcaat tattaatctt ttgtttcctc
      361 gtcattgttc tcgttccctt tcttccttgt ttctttttct gcacaatatt tcaagctata
      421 ccaagcatac aatcaactcc aagctttgca aagatggata aagcggaatt aattcccgag
      481 cctccaaaaa agaagagaaa ggtcgaattg ggtaccgccg ccaattttaa tcaaagtggg
      541 aatattgctg atagctcatt gtccttcact ttcactaaca gtagcaacgg tccgaacctc
      601 ataacaactc aaacaaattc tcaagcgctt tcacaaccaa ttgcctcctc taacgttcat
      661 gataacttca tgaataatga aatcacggct agtaaaattg atgatggtaa taattcaaaa
      721 ccactgtcac ctggttggac ggaccaaact gcgtataacg cgtttggaat cactacaggg
      781 atgtttaata ccactacaat ggatgatgta tataactatc tattcgatga tgaagatacc
      841 ccaccaaacc caaaaaaaga gatctctatg gcttacccat acgatgttcc agattacgct
      901 agcttgggtg gtcatatggc catggaggcc ccggggatcc gaatgattca ggcgattctg
      961 gttttcaaca accatgggaa gccacggcta gtccgcttct accagcgttt cccagaagaa
     1021 attcaacagc agattgttcg agagactttc catctagtcc tcaagcggga tgacaacatc
     1081 tgtaacttct tggagggtgg aagtttgatt ggtggctctg actacaaact gatctaccgg
     1141 cactatgcta ccctctactt tgtattttgt gtggattcct cagagagtga acttggaatc
     1201 ttggacctca tccaggtttt tgtggaaact ctggataagt gtttcgaaaa tgtgtgtgaa
     1261 ttggatttga tcttccatat ggataaggtg cactacatcc tccaggaggt ggtgatgggt
     1321 gggatggtgt tggaaacaaa catgaatgaa atcgtggctc agattgaggc tcaaaacagg
     1381 ctggagaaat ccgagggtgg cctttcagca gcccctgcgc gggctgtgtc tgctgtgaaa
     1441 aacatcaacc tgccagagat tcctcggaac atcaacattg gcgatctcaa catcaaagtt
     1501 cccaacctgt cccagtttgt ctgagctcga gagatctatg aatcgtagat actgaaaaac
     1561 cccgcaagtt cacttcaact gtgcatcgtg caccatctca atttctttca tttatacatc
     1621 gttttgcctt cttttatgta actatactcc tctaagtttc aatcttggcc atgtaacctc
     1681 tgatctatag aattttttaa atgactagaa ttaatgccca tctttttttt ggacctaaat
     1741 tcttcatgaa aatatattac gagggcttat tcagaagctt tggacttctt cgccagaggt
     1801 ttggtcaagt ctccaatcaa ggttgtcggc ttgtctacct tgccagaaat ttacgaaaag
     1861 atggaaaagg gtcaaatcgt tggtagatac gttgttgaca cttctaaata agcgaatttc
     1921 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     1981 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     2041 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc
     2101 atgccggtcg agggacctaa taacttcgta tagcatacat tatacgaagt tatattaagg
     2161 gttccggatc gcggccgctc gacctgcagc caagctagct tggctggacg taaactcctc
     2221 ttcagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttattgaa
     2281 tatgatcgga attggtcgag ggacctaata acttcgtata gcatacatta tacgaagtta
     2341 tattaagggt tccggatcgc ggccgctcga cctgcagcca agctagcttg gctggacgta
     2401 aactcctctt cagacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg
     2461 ttattgaata tgatcggaat tggtcgacta cgtcgttaag gccgtttctg acagagtaaa
     2521 attcttgagg gaactttcac cattatggga aatggttcaa gaaggtattg acttaaactc
     2581 catcaaatgg tcaggtcatt gagtgttttt tatttgttgt attttttttt ttttagagaa
     2641 aatcctccaa tatataaatt aggaatcata gtttcatgat tttctgttac acctaacttt
     2701 ttgtgtggtg ccctcctcct tgtcaatatt aatgttaaag tgcaattctt tttccttatc
     2761 acgttgagcc attagtatca atttgcttac ctgtattcct ttacatcctc ctttttctcc
     2821 ttcttgataa atgtatgtag attgcgtata tagtttcgtc taccctatga acatattcca
     2881 ttttgtaatt tcgtgtcgtt tctattatga atttcattta taaagtttat gtacaaatat
     2941 cataaaaaaa gagaatcttt ttaagcaagg attttcttaa cttcttcggc gacagcatca
     3001 ccgacttcgg tggtactgtt ggaaccacct aaatcaccag ttctgatacc tgcatccaaa
     3061 acctttttaa ctgcatcttc aatggcctta ccttcttcag gcaagttcaa tgacaatttc
     3121 aacatcattg cagcagacaa gatagtggcg atagggttga ccttattctt tggcaaatct
     3181 ggagcagaac cgtggcatgg ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag
     3241 gccaaggacg cagatggcaa caaacccaag gaacctggga taacggaggc ttcatcggag
     3301 atgatatcac caaacatgtt gctggtgatt ataataccat ttaggtgggt tgggttctta
     3361 actaggatca tggcggcaga atcaatcaat tgatgttgaa ccttcaatgt aggaaattcg
     3421 ttcttgatgg tttcctccac agtttttctc cataatcttg aagaggccaa aacattagct
     3481 ttatccaagg accaaatagg caatggtggc tcatgttgta gggccatgaa agcggccatt
     3541 cttgtgattc tttgcacttc tggaacggtg tattgttcac tatcccaagc gacaccatca
     3601 ccatcgtctt cctttctctt accaaagtaa atacctccca ctaattctct gacaacaacg
     3661 aagtcagtac ctttagcaaa ttgtggcttg attggagata agtctaaaag agagtcggat
     3721 gcaaagttac atggtcttaa gttggcgtac aattgaagtt ctttacggat ttttagtaaa
     3781 ccttgttcag gtctaacact acctgtaccc catttaggac cacccacagc acctaacaaa
     3841 acggcatcag ccttcttgga ggcttccagc gcctcatctg gaagtggaac acctgtagca
     3901 tcgatagcag caccaccaat taaatgattt tcgaaatcga acttgacatt ggaacgaaca
     3961 tcagaaatag ctttaagaac cttaatggct tcggctgtga tttcttgacc aacgtggtca
     4021 cctggcaaaa cgacgatctt cttaggggca gacattagaa tggtatatcc ttgaaatata
     4081 tatatatatt gctgaaatgt aaaaggtaag aaaagttaga aagtaagacg attgctaacc
     4141 acctattgga aaaaacaata ggtccttaaa taatattgtc aacttcaagt attgtgatgc
     4201 aagcatttag tcatgaacgc ttctctattc tatatgaaaa gccggttccg gcgctctcac
     4261 ctttcctttt tctcccaatt tttcagttga aaaaggtata tgcgtcaggc gacctctgaa
     4321 attaacaaaa aatttccagt catcgaattt gattctgtgc gatagcgccc ctgtgtgttc
     4381 tcgttatgtt gaggaaaaaa ataatggttg ctaagagatt cgaactcttg catcttacga
     4441 tacctgagta ttcccacagt tccttaccac tcttttgtta ctctattgat ccagctcagc
     4501 aaaggcagtg tgatctaaga ttctatcttc gcgatgtagt aaaactagct agaccgagaa
     4561 agagactaga aatgcaaaag gcacttctac aatggctgcc atcattatta tccgatgtga
     4621 cgctgcagct tctcaatgat attcgaatac gctttgagga gatacagcct aatatccgac
     4681 aaactgtttt acagatttac gatcgtactt gttacccatc attgaatttt gaacatccga
     4741 acctgggagt tttccctgaa acagatagta tatttgaacc tgtataataa tatatagtct
     4801 agcgctttac ggaagacaat gtatgtattt cggttcctgg agaaactatt gcatctattg
     4861 cataggtaat cttgcacgtc gcatccccgg ttcattttct gcgtttccat cttgcacttc
     4921 aatagcatat ctttgttaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     4981 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     5041 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     5101 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     5161 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     5221 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     5281 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     5341 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     5401 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     5461 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     5521 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     5581 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     5641 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     5701 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     5761 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     5821 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt
     5881 ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata
     5941 ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca
     6001 tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg
     6061 agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt
     6121 aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta
     6181 tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag
     6241 tctcatcctt caatgctatc atttcctttg atattggatc atatgcatag taccgagaaa
     6301 ctagtgcgaa gtagtgatca ggtattgctg ttatctgatg agtatacgtt gtcctggcca
     6361 cggcagaagc acgcttatcg ctccaatttc ccacaacatt agtcaactcc gttaggccct
     6421 tcattgaaag aaatgaggtc atcaaatgtc ttccaatgtg agattttggg ccatttttta
     6481 tagcaaagat tgaataaggc gcatttttct tcaaagctca ggtggcactt ttcggggaaa
     6541 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6601 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6661 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6721 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6781 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6841 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     6901 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6961 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7021 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7081 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7141 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     7201 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7261 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7321 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7381 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7441 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7501 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7561 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7621 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7681 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7741 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7801 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     7861 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7921 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7981 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8041 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8101 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8161 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8221 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8281 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8341 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8401 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     8461 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     8521 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     8581 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     8641 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgt
//
