LOCUS       sequence-387637-        8418 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of GAL4 transcriptional activation domain (AD)-AP-1 
            sigma-1C fusion protein in yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8418)
  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 8418)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pGADT7-AP-1 sigma-1C
FEATURES             Location/Qualifiers
     source          1..8418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        763..1467
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1513..1533
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1549..1890
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     primer_bind     1896..1915
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1896..1914
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1921..1930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1933..1959
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      2838..3025
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2893..2917)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3142..4236)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4237..4642)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4684..4700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4684..4700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4697..4719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4708..4724
                     /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(4732..4738,4739..4756,4757..4762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    4853..4886
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4964..4997
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5066..5083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5237..5825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5317..5336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5996..6787,6788..6856))
                     /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     6619..6638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6857..6961)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7029..7047
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7085..7107)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7207..7226
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7243..8407
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgcaggtcga gatccgggat cgaagaaatg atggtaaatg aaataggaaa tcaaggagca
       61 tgaaggcaaa agacaaatat aagggtcgaa cgaaaaataa agtgaaaagt gttgatatga
      121 tgtatttggc tttgcggcgc cgaaaaaacg agtttacgca attgcacaat catgctgact
      181 ctgtggcgga cccgcgctct tgccggcccg gcgataacgc tgggcgtgag gctgtgcccg
      241 gcggagtttt ttgcgcctgc attttccaag gtttaccctg cgctaagggg cgagattgga
      301 gaagcaataa gaatgccggt tggggttgcg atgatgacga ccacgacaac tggtgtcatt
      361 atttaagttg ccgaaagaac ctgagtgcat ttgcaacatg agtatactag aagaatgagc
      421 caagacttgc gagacgcgag tttgccggtg gtgcgaacaa tagagcgacc atgaccttga
      481 aggtgagacg cgcataaccg ctagagtact ttgaagagga aacagcaata gggttgctac
      541 cagtataaat agacaggtac atacaacact ggaaatggtt gtctgtttga gtacgctttc
      601 aattcatttg ggtgtgcact ttattatgtt acaatatgga agggaacttt acacttctcc
      661 tatgcacata tattaattaa agtccaatgc tagtagagaa ggggggtaac acccctccgc
      721 gctcttttcc gatttttttc taaaccgtgg aatatttcgg atatcctttt gttgtttccg
      781 ggtgtacaat atggacttcc tcttttctgg caaccaaacc catacatcgg gattcctata
      841 ataccttcgt tggtctccct aacatgtagg tggcggaggg gagatataca atagaacaga
      901 taccagacaa gacataatgg gctaaacaag actacaccaa ttacactgcc tcattgatgg
      961 tggtacataa cgaactaata ctgtagccct agacttgata gccatcatca tatcgaagtt
     1021 tcactaccct ttttccattt gccatctatt gaagtaataa taggcgcatg caacttcttt
     1081 tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca atgacaaaaa
     1141 aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac agatgtcgtt
     1201 gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc cttcattcac
     1261 gcacactact ctctaatgag caacggtata cggccttcct tccagttact tgaatttgaa
     1321 ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt attaatcttt
     1381 tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg cacaatattt
     1441 caagctatac caagcataca atcaactcca agctttgcaa agatggataa agcggaatta
     1501 attcccgagc ctccaaaaaa gaagagaaag gtcgaattgg gtaccgccgc caattttaat
     1561 caaagtggga atattgctga tagctcattg tccttcactt tcactaacag tagcaacggt
     1621 ccgaacctca taacaactca aacaaattct caagcgcttt cacaaccaat tgcctcctct
     1681 aacgttcatg ataacttcat gaataatgaa atcacggcta gtaaaattga tgatggtaat
     1741 aattcaaaac cactgtcacc tggttggacg gaccaaactg cgtataacgc gtttggaatc
     1801 actacaggga tgtttaatac cactacaatg gatgatgtat ataactatct attcgatgat
     1861 gaagataccc caccaaaccc aaaaaaagag atctttaata cgactcacta tagggcgagc
     1921 gccgccatgg agtacccata cgacgtacca gattacgctc atatggccat ggaggccagt
     1981 gaattcatga tacatttcat attgctcttc agtcgacaag ggaaattacg gctacagaaa
     2041 tggtacatca ctctccctga taaagagagg aagaagatca cccgggaaat tgttcagatt
     2101 attctctccc gtggtcacag gacaagcagt tttgttgact ggaaggagct aaaacttgtt
     2161 tataaaaggt atgctagttt atatttttgc tgtgcaatag aaaatcagga caatgagctc
     2221 ttgacgctag agattgtgca tcgttacgtg gagctgctgg acaaatattt tggaaatgtc
     2281 tgtgagctgg atattatctt taattttgaa aaggcttatt tcatcctgga cgagtttata
     2341 ataggtgggg aaattcagga aacatccaag aaaattgctg tcaaagccat tgaagactct
     2401 gatatgttac aggaggtcag tacggtttgc caaaccatgg gagaaagatg actcgagctg
     2461 cagatgaatc gtagatactg aaaaaccccg caagttcact tcaactgtgc atcgtgcacc
     2521 atctcaattt ctttcattta tacatcgttt tgccttcttt tatgtaacta tactcctcta
     2581 agtttcaatc ttggccatgt aacctctgat ctatagaatt ttttaaatga ctagaattaa
     2641 tgcccatctt ttttttggac ctaaattctt catgaaaata tattacgagg gcttattcag
     2701 aagctttgga cttcttcgcc agaggtttgg tcaagtctcc aatcaaggtt gtcggcttgt
     2761 ctaccttgcc agaaatttac gaaaagatgg aaaagggtca aatcgttggt agatacgttg
     2821 ttgacacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     2881 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     2941 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg
     3001 ctcttattga ccacacctct accggccggt cgaaattccc ctaccctatg aacatattcc
     3061 attttgtaat ttcgtgtcgt ttctattatg aatttcattt ataaagttta tgtacaaata
     3121 tcataaaaaa agagaatctt tttaagcaag gattttctta acttcttcgg cgacagcatc
     3181 accgacttcg gtggtactgt tggaaccacc taaatcacca gttctgatac ctgcatccaa
     3241 aaccttttta actgcatctt caatggcctt accttcttca ggcaagttca atgacaattt
     3301 caacatcatt gcagcagaca agatagtggc gatagggttg accttattct ttggcaaatc
     3361 tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt ctggcaaaga
     3421 ggccaaggac gcagatggca acaaacccaa ggaacctggg ataacggagg cttcatcgga
     3481 gatgatatca ccaaacatgt tgctggtgat tataatacca tttaggtggg ttgggttctt
     3541 aactaggatc atggcggcag aatcaatcaa ttgatgttga accttcaatg taggaaattc
     3601 gttcttgatg gtttcctcca cagtttttct ccataatctt gaagaggcca aaacattagc
     3661 tttatccaag gaccaaatag gcaatggtgg ctcatgttgt agggccatga aagcggccat
     3721 tcttgtgatt ctttgcactt ctggaacggt gtattgttca ctatcccaag cgacaccatc
     3781 accatcgtct tcctttctct taccaaagta aatacctccc actaattctc tgacaacaac
     3841 gaagtcagta cctttagcaa attgtggctt gattggagat aagtctaaaa gagagtcgga
     3901 tgcaaagtta catggtctta agttggcgta caattgaagt tctttacgga tttttagtaa
     3961 accttgttca ggtctaacac tacctgtacc ccatttagga ccacccacag cacctaacaa
     4021 aacggcatca gccttcttgg aggcttccag cgcctcatct ggaagtggga cacctgtagc
     4081 ttcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat tggaacgaac
     4141 atcagaaata gctttaagaa ccttaatggc ttcggctgtg atttcttgac caacgtggtc
     4201 acctggcaaa acgacgatct tcttaggggc agacattaga atggtatatc cttgaaatat
     4261 atatatatat tgctgaaatg taaaaggtaa gaaaagttag aaagtaagac gattgctaac
     4321 cacctattgg aaaaaacaat aggtccttaa ataatattgt caacttcaag tattgtgatg
     4381 caagcattta gtcatgaacg cttctctatt ctatatgaaa agccggttcc ggcgctctca
     4441 cctttccttt ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg cgacctctga
     4501 aattaacaaa aaatttccag tcatcgaatt tgattctgtg cgatagcgcc cctgtgtgtt
     4561 ctcgttatgt tgaggaaaaa aataatggtt gctaagagat tcgaactctt gcatcttacg
     4621 atacctgagt attcccacag ttgggggatc tcgactctag ctagaggatc aattcgtaat
     4681 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgataacttc
     4861 gtataatgta tgctatacga agttattagg tctgaagagg agtttacgtc cagccaagct
     4921 agcttggctg caggtcgagc ggccgcgatc cggaaccctt aatataactt cgtataatgt
     4981 atgctatacg aagttatcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5041 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5101 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5161 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5221 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5281 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5341 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5401 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5461 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5521 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5581 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5641 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5701 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5761 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5821 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5881 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5941 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6001 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6061 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6121 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6181 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6241 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6301 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6361 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6421 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6481 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6541 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6601 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6661 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6721 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6781 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6841 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6901 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6961 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     7021 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     7081 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     7141 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     7201 actatgcggc atcagagcag attgtactga gagtgcacca taacgcattt aagcataaac
     7261 acgcactatg ccgttcttct catgtatata tatatacagg caacacgcag atataggtgc
     7321 gacgtgaaca gtgagctgta tgtgcgcagc tcgcgttgca ttttcggaag cgctcgtttt
     7381 cggaaacgct ttgaagttcc tattccgaag ttcctattct ctagctagaa agtataggaa
     7441 cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg cactttcaaa aaaccaaaaa
     7501 cgcaccggac tgtaacgagc tactaaaata ttgcgaatac cgcttccaca aacattgctc
     7561 aaaagtatct ctttgctata tatctctgtg ctatatccct atataaccta cccatccacc
     7621 tttcgctcct tgaacttgca tctaaactcg acctctacat tttttatgtt tatctctagt
     7681 attactcttt agacaaaaaa attgtagtaa gaactattca tagagtgaat cgaaaacaat
     7741 acgaaaatgt aaacatttcc tatacgtagt atatagagac aaaatagaag aaaccgttca
     7801 taattttctg accaatgaag aatcatcaac gctatcactt tctgttcaca aagtatgcgc
     7861 aatccacatc ggtatagaat ataatcgggg atgcctttat cttgaaaaaa tgcacccgca
     7921 gcttcgctag taatcagtaa acgcgggaag tggagtcagg ctttttttat ggaagagaaa
     7981 atagacacca aagtagcctt cttctaacct taacggacct acagtgcaaa aagttatcaa
     8041 gagactgcat tatagagcgc acaaaggaga aaaaaagtaa tctaagatgc tttgttagaa
     8101 aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat tctttgttgg
     8161 taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt
     8221 tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca cagattcttc
     8281 gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct
     8341 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga agcacagatg
     8401 cttcgttgct tgcatgcc
//
