LOCUS       sequence-388878-        7458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 1) GAL4 DNA-binding domain (BD)-tyrosinase cytosolic 
            tail fusion protein, 2) HA epitope and nuclear localization signal 
            (NLS) AP-1 sigma-1A fusion protein in yeast (yeast three-hybrid 
            assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7458)
  AUTHORS   Mattera R, Boehm M, Chaudhuri R, Prabhu Y, Bonifacino JS
  TITLE     Conservation and diversification of dileucine signal recognition by 
            adaptor protein (AP) complex variants.
  JOURNAL   J Biol Chem. 2011 Jan 21;286(3):2022-30. doi: 
            10.1074/jbc.M110.197178. Epub 2010 Nov 19.
  PUBMED    21097499
REFERENCE   2  (bases 1 to 7458)
  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: 
            "10.1074/jbc.M110.197178"; journalName: "J Biol Chem"; date: 
            "2011-01-21- 21"; volume: "286"; issue: "3"; pages: "2022-30"
COMMENT     Sequence Label: pBridge-tyrosinase.sigma-1A
FEATURES             Location/Qualifiers
     source          1..7458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1992..2010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2048..2070)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2170..2189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      2206..3370
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3375..3776
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3804..4244
                     /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      4712..4899
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4767..4791)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4952..5626)
                     /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(5627..5728)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5742..5758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5742..5758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5755..5777)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5766..5782
                     /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(5790..5796,5797..5814,5815..5820))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5835..5856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6050..6400
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             6422..6448
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6428..6454
                     /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"
     CDS             6470..6490
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7126..7396
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1981 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     2041 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2101 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     2161 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataacgca tttaagcata
     2221 aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg
     2281 tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt
     2341 tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagcta gaaagtatag
     2401 gaacttcaga gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa
     2461 aaacgcaccg gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg
     2521 ctcaaaagta tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc
     2581 acctttcgct ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct
     2641 agtattactc tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac
     2701 aatacgaaaa tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt
     2761 tcataatttt ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg
     2821 cgcaatccac atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc
     2881 gcagcttcgc tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag
     2941 aaaatagaca ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat
     3001 caagagactg cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta
     3061 gaaaaatagc gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt
     3121 tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt
     3181 gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc
     3241 ttcgttggta aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat
     3301 tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag
     3361 atgcttcgtt gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg
     3421 ttgtctcacc atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa
     3481 cgacaaagac agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc
     3541 acacgaaact ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac
     3601 ggccttcctt ccagttactt gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag
     3661 tataaataga cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc
     3721 ttccttgttt ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa
     3781 gcttgaagca agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata
     3841 tttgccgact taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga
     3901 agaacaactg ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac
     3961 atctgacaga agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc
     4021 ctcgagaaga ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt
     4081 taacaggatt atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt
     4141 cagtggagac tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat
     4201 cggaagagag tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcccgggga
     4261 tcctatgtct tcagaagaag aagaagaaga agcaacccca ggaggaaagg cagccactcc
     4321 tcatggacaa agacgactac cacagcttgc tgtatcagag ccatctgtga acatcctagg
     4381 aaacagagtg ggactgaaag gttttacctc actcgaccta tttgttggtg tttctacaaa
     4441 tttaaactag tataaaacat agaccatagc tgtttggctt tttttcagac ccatgttttt
     4501 tcctaagtcc tagtttctaa gaaatgactg ggatttgcta aaatatatat atatataaat
     4561 aataacttac taatagctaa ataaaatttc ctcttacaac taattgagct ggtttttatg
     4621 aatgtgtctt aattatttaa acttgaggca catttttgtt ttccttactt cattgtgaat
     4681 ttccggaatt ctgcagccaa gctaattccg ggcgaatttc ttatgattta tgatttttat
     4741 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
     4801 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
     4861 atagcatgag gtcgctctta ttgaccacac ctctaccggc atgccggcaa gtgcacaaac
     4921 aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg acgaaatttg
     4981 ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa
     5041 taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca
     5101 taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat
     5161 ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt
     5221 tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa
     5281 ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc agttggacga
     5341 tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga ttacgaaaca
     5401 cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca agacttgaaa
     5461 ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat ataataccca
     5521 gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc aagccgcaaa
     5581 ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatactc caagctgcct
     5641 ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca ccaatgccct ccctcttggc
     5701 cctctccttt tcttttttcg accgaattaa ttcgtaatca tgtcatagct gtttcctgtg
     5761 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5821 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5881 ttccagtcgg gaaacctgtc gtgccaggaa gatccgaggc ctagcttcta attcttccaa
     5941 catacaatgg gagtttggcc gagtggttta aggcgtcaga tttaggtgga tttaacctct
     6001 aaaatctctg atatcttcgg atgcaagggt tcgaatccct tagctctcat tattttttgc
     6061 tttttctctt gaggtcacat gatcgcaaaa tggcaaatgg cacgtgaagc tgtcgatatt
     6121 ggggaactgt ggtggttggc aaatgactaa ttaagttagt caaggcgcca tcctcatgaa
     6181 aactgtgtaa cataataacc gaagtgtcga aaaggtggca ccttgtccaa ttgaacacgc
     6241 tcgatgaaaa aaataagata tatataaggt taagtaaagc gtctgttaga aaggaagttt
     6301 ttcctttttc ttgctctctt gtcttttcat ctactatttc cttcgtgtaa tacagggtcg
     6361 tcagatacat agatacaatt ctattacccc catccataca atgggccata tggcttctag
     6421 ctatccttat gacgtgcctg actatgccag cctgggagga ccttctagtc ctaagaagaa
     6481 gagaaaggtg gcggccgcca gtgtgatgga tatctgtaga attcgccctt ggacggccgc
     6541 catgatgcgg ttcatgctat tattcagccg gcggggaaaa ctgcggctgc aaaaatggta
     6601 cctggccact tcggacaagg aacggaagaa gatggtgcgc gagctcatgc aggttgtcct
     6661 ggctcgaaag cccaagatgt gcagcttcct ggagtggagg gacctcaaag ttgtctataa
     6721 gagatatgcc agcctctact tctgctgcgc catcgagggc caagacaatg agctcatcac
     6781 actggagctg atccaccgat acgtggagct cttagacaaa tactttggca gtgtgtgcga
     6841 gctggacatc atcttcaact ttgagaaggc ctacttcatc ctggatgagt ttttgatggg
     6901 gggggatgtc caggacacct ccaagaagag tgtgctgaaa gccatcgagc aggctgacct
     6961 actgcaagag gaggatgagt cgccacggag tgtgctggag gagatgggtt tggcatagag
     7021 atcttcgggc tgatctccca tgtctctact ggtggtggtg cttctttgga attattggaa
     7081 ggtaaggaat tgccaggtgt tgctttctta tccgaaaaga aataaattga attgaattga
     7141 aatcgataga tcaatttttt tcttttctct ttccccatcc tttacgctaa aataatagtt
     7201 tattttattt tttgaatatt ttttatttat atacgtatat atagactatt atttatcttt
     7261 taatgattat taagattttt attaaaaaaa aattcgctcc tcttttaatg cctttatgca
     7321 gttttttttt cccattcgat atttctatgt tcgggttcag cgtattttaa gtttaataac
     7381 tcgaaaattc tgcgttcgtt aaagctaggc ctcggatctt cctgcattaa tgaatcggcc
     7441 aacgcgcggg gagaggcg
//
