LOCUS       sequence-387539-        8885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 transcriptional activation domain (AD)-AP-3 
            sigma-3A fusion protein in yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8885)
  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 8885)
  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-3A
FEATURES             Location/Qualifiers
     source          1..8885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..27
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        56..457
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             503..523
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             539..880
                     /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             893..919
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             899..925
                     /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      1933..2120
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1988..2012)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(2141..2174)
                     /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(2252..2285)
                     /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(2329..2362)
                     /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(2440..2473)
                     /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(2517..2550)
                     /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(2628..2661)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3170..4264)
                     /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(4265..4670)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      5147..6489
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6110..6157)
                     /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        6754..6858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6859..6927,6928..7719)
                     /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(7077..7096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7890..8478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8379..8398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8632..8649
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8664..8804
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8730..8749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
       61 caacttcttt tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca
      121 atgacaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac
      181 agatgtcgtt gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc
      241 cttcattcac gcacactact ctctaatgag caacggtata cggccttcct tccagttact
      301 tgaatttgaa ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt
      361 attaatcttt tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg
      421 cacaatattt caagctatac caagcataca atcaactcca agctttgcaa agatggataa
      481 agcggaatta attcccgagc ctccaaaaaa gaagagaaag gtcgaattgg gtaccgccgc
      541 caattttaat caaagtggga atattgctga tagctcattg tccttcactt tcactaacag
      601 tagcaacggt ccgaacctca taacaactca aacaaattct caagcgcttt cacaaccaat
      661 tgcctcctct aacgttcatg ataacttcat gaataatgaa atcacggcta gtaaaattga
      721 tgatggtaat aattcaaaac cactgtcacc tggttggacg gaccaaactg cgtataacgc
      781 gtttggaatc actacaggga tgtttaatac cactacaatg gatgatgtat ataactatct
      841 attcgatgat gaagataccc caccaaaccc aaaaaaagag atctctatgg cttacccata
      901 cgatgttcca gattacgcta gcttgggtgg tcatatggcc atggaggccc cggggatccg
      961 agccatgatc aaggcgatcc taatcttcaa caaccacggg aagccgcggc tctccaagtt
     1021 ctaccagccc tacagtgaag atacacaaca gcaaatcatc agggagactt tccatttggt
     1081 atctaagaga gatgaaaatg tttgtaattt cctagaagga ggattattaa ttggaggatc
     1141 tgacaacaaa ctgatttata gacattatgc aacgttatat tttgtcttct gtgtggattc
     1201 ttcagaaagt gaacttggca ttttagatct aattcaagta tttgtggaaa cattagacaa
     1261 atgttttgaa aatgtctgtg agctggattt gattttccat gtagacaagg ttcacaatat
     1321 tcttgcagaa atggtgatgg ggggaatggt attggagaca aatatgaatg agattgttac
     1381 acaaattgat gcacaaaata agctggaaaa atctgaggct ggcttagcag gagctccagc
     1441 ccgtgctgta tcagctgtaa agaatatgaa tcttcctgag atcccaagaa atattaacat
     1501 tggtgacatc agtataaaag tgccaaacct gccctctttt aaataactcg agagatctat
     1561 gaatcgtaga tactgaaaaa ccccgcaagt tcacttcaac tgtgcatcgt gcaccatctc
     1621 aatttctttc atttatacat cgttttgcct tcttttatgt aactatactc ctctaagttt
     1681 caatcttggc catgtaacct ctgatctata gaatttttta aatgactaga attaatgccc
     1741 atcttttttt tggacctaaa ttcttcatga aaatatatta cgagggctta ttcagaagct
     1801 ttggacttct tcgccagagg tttggtcaag tctccaatca aggttgtcgg cttgtctacc
     1861 ttgccagaaa tttacgaaaa gatggaaaag ggtcaaatcg ttggtagata cgttgttgac
     1921 acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     1981 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     2041 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt
     2101 attgaccaca cctctaccgg catgccggtc gagggaccta ataacttcgt atagcataca
     2161 ttatacgaag ttatattaag ggttccggat cgcggccgct cgacctgcag ccaagctagc
     2221 ttggctggac gtaaactcct cttcagacct aataacttcg tatagcatac attatacgaa
     2281 gttatattaa gggttattga atatgatcgg aattggtcga gggacctaat aacttcgtat
     2341 agcatacatt atacgaagtt atattaaggg ttccggatcg cggccgctcg acctgcagcc
     2401 aagctagctt ggctggacgt aaactcctct tcagacctaa taacttcgta tagcatacat
     2461 tatacgaagt tatattaagg gttattgaat atgatcggaa ttggtcgagg gacctaataa
     2521 cttcgtatag catacattat acgaagttat attaagggtt ccggatcgcg gccgctcgac
     2581 ctgcagccaa gctagcttgg ctggacgtaa actcctcttc agacctaata acttcgtata
     2641 gcatacatta tacgaagtta tattaagggt tattgaatat gatcggaatt ggtcgactac
     2701 gtcgttaagg ccgtttctga cagagtaaaa ttcttgaggg aactttcacc attatgggaa
     2761 atggttcaag aaggtattga cttaaactcc atcaaatggt caggtcattg agtgtttttt
     2821 atttgttgta tttttttttt tttagagaaa atcctccaat atataaatta ggaatcatag
     2881 tttcatgatt ttctgttaca cctaactttt tgtgtggtgc cctcctcctt gtcaatatta
     2941 atgttaaagt gcaattcttt ttccttatca cgttgagcca ttagtatcaa tttgcttacc
     3001 tgtattcctt tacatcctcc tttttctcct tcttgataaa tgtatgtaga ttgcgtatat
     3061 agtttcgtct accctatgaa catattccat tttgtaattt cgtgtcgttt ctattatgaa
     3121 tttcatttat aaagtttatg tacaaatatc ataaaaaaag agaatctttt taagcaagga
     3181 ttttcttaac ttcttcggcg acagcatcac cgacttcggt ggtactgttg gaaccaccta
     3241 aatcaccagt tctgatacct gcatccaaaa cctttttaac tgcatcttca atggccttac
     3301 cttcttcagg caagttcaat gacaatttca acatcattgc agcagacaag atagtggcga
     3361 tagggttgac cttattcttt ggcaaatctg gagcagaacc gtggcatggt tcgtacaaac
     3421 caaatgcggt gttcttgtct ggcaaagagg ccaaggacgc agatggcaac aaacccaagg
     3481 aacctgggat aacggaggct tcatcggaga tgatatcacc aaacatgttg ctggtgatta
     3541 taataccatt taggtgggtt gggttcttaa ctaggatcat ggcggcagaa tcaatcaatt
     3601 gatgttgaac cttcaatgta ggaaattcgt tcttgatggt ttcctccaca gtttttctcc
     3661 ataatcttga agaggccaaa acattagctt tatccaagga ccaaataggc aatggtggct
     3721 catgttgtag ggccatgaaa gcggccattc ttgtgattct ttgcacttct ggaacggtgt
     3781 attgttcact atcccaagcg acaccatcac catcgtcttc ctttctctta ccaaagtaaa
     3841 tacctcccac taattctctg acaacaacga agtcagtacc tttagcaaat tgtggcttga
     3901 ttggagataa gtctaaaaga gagtcggatg caaagttaca tggtcttaag ttggcgtaca
     3961 attgaagttc tttacggatt tttagtaaac cttgttcagg tctaacacta cctgtacccc
     4021 atttaggacc acccacagca cctaacaaaa cggcatcagc cttcttggag gcttccagcg
     4081 cctcatctgg aagtggaaca cctgtagcat cgatagcagc accaccaatt aaatgatttt
     4141 cgaaatcgaa cttgacattg gaacgaacat cagaaatagc tttaagaacc ttaatggctt
     4201 cggctgtgat ttcttgacca acgtggtcac ctggcaaaac gacgatcttc ttaggggcag
     4261 acattagaat ggtatatcct tgaaatatat atatatattg ctgaaatgta aaaggtaaga
     4321 aaagttagaa agtaagacga ttgctaacca cctattggaa aaaacaatag gtccttaaat
     4381 aatattgtca acttcaagta ttgtgatgca agcatttagt catgaacgct tctctattct
     4441 atatgaaaag ccggttccgg cgctctcacc tttccttttt ctcccaattt ttcagttgaa
     4501 aaaggtatat gcgtcaggcg acctctgaaa ttaacaaaaa atttccagtc atcgaatttg
     4561 attctgtgcg atagcgcccc tgtgtgttct cgttatgttg aggaaaaaaa taatggttgc
     4621 taagagattc gaactcttgc atcttacgat acctgagtat tcccacagtt ccttaccact
     4681 cttttgttac tctattgatc cagctcagca aaggcagtgt gatctaagat tctatcttcg
     4741 cgatgtagta aaactagcta gaccgagaaa gagactagaa atgcaaaagg cacttctaca
     4801 atggctgcca tcattattat ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg
     4861 ctttgaggag atacagccta atatccgaca aactgtttta cagatttacg atcgtacttg
     4921 ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa cagatagtat
     4981 atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg tatgtatttc
     5041 ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg catccccggt
     5101 tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg aagcatctgt
     5161 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     5221 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     5281 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     5341 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     5401 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     5461 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     5521 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     5581 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     5641 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     5701 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     5761 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     5821 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     5881 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     5941 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     6001 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     6061 ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     6121 actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc
     6181 ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc
     6241 tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta
     6301 aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga
     6361 tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc
     6421 tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga
     6481 tattggatca tatgcatagt accgagaaac tagtgcgaag tagtgatcag gtattgctgt
     6541 tatctgatga gtatacgttg tcctggccac ggcagaagca cgcttatcgc tccaatttcc
     6601 cacaacatta gtcaactccg ttaggccctt cattgaaaga aatgaggtca tcaaatgtct
     6661 tccaatgtga gattttgggc cattttttat agcaaagatt gaataaggcg catttttctt
     6721 caaagctcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6781 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6841 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6901 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6961 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7021 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7081 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     7141 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7201 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7261 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7321 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7381 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     7441 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7501 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7561 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7621 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7681 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7741 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7801 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     8041 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     8221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8521 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8581 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     8641 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     8701 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     8761 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     8821 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     8881 tgacc
//
