LOCUS       sequence-388893-        7368 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-2 sigma-2 fusion protein in yeast (yeast three-hybrid 
            assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  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 7368)
  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-2
FEATURES             Location/Qualifiers
     source          1..7368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1167..1568
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1596..2036
                     /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      2504..2691
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2559..2583)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2744..3418)
                     /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(3419..3520)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3534..3550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3534..3550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3547..3569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3558..3574
                     /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(3582..3588,3589..3606,3607..3612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3627..3648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3842..4192
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             4214..4240
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4220..4246
                     /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             4262..4282
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4828..5098
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(5189..5206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5360..5948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5440..5459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6119..6910,6911..6979))
                     /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     6742..6761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6980..7084)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7152..7170
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7208..7230)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7330..7349
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(7366..7368,1..1162)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 catttaagca taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca
       61 cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc
      121 ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctagc
      181 tagaaagtat aggaacttca gagcgctttt gaaaaccaaa agcgctctga agacgcactt
      241 tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta aaatattgcg aataccgctt
      301 ccacaaacat tgctcaaaag tatctctttg ctatatatct ctgtgctata tccctatata
      361 acctacccat ccacctttcg ctccttgaac ttgcatctaa actcgacctc tacatttttt
      421 atgtttatct ctagtattac tctttagaca aaaaaattgt agtaagaact attcatagag
      481 tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac gtagtatata gagacaaaat
      541 agaagaaacc gttcataatt ttctgaccaa tgaagaatca tcaacgctat cactttctgt
      601 tcacaaagta tgcgcaatcc acatcggtat agaatataat cggggatgcc tttatcttga
      661 aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag tcaggctttt
      721 tttatggaag agaaaataga caccaaagta gccttcttct aaccttaacg gacctacagt
      781 gcaaaaagtt atcaagagac tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa
      841 gatgctttgt tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta
      901 tagattcttt gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag
      961 ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt tacaaaaatg
     1021 aagcacagat tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa
     1081 tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa
     1141 aatgaagcac agatgcttcg ttgcttgcat gcaacttctt ttcttttttt ttcttttctc
     1201 tctcccccgt tgttgtctca ccatatccgc aatgacaaaa aaatgatgga agacactaaa
     1261 ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg
     1321 gtatctcgaa gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga
     1381 gcaacggtat acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt
     1441 cttgctatca agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc
     1501 tcgttccctt tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac
     1561 aatcaactcc aagcttgaag caagcctcct gaaagatgaa gctactgtct tctatcgaac
     1621 aagcatgcga tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa ccgaagtgcg
     1681 ccaagtgtct gaagaacaac tgggagtgtc gctactctcc caaaaccaaa aggtctccgc
     1741 tgactagggc acatctgaca gaagtggaat caaggctaga aagactggaa cagctatttc
     1801 tactgatttt tcctcgagaa gaccttgaca tgattttgaa aatggattct ttacaggata
     1861 taaaagcatt gttaacagga ttatttgtac aagataatgt gaataaagat gccgtcacag
     1921 atagattggc ttcagtggag actgatatgc ctctaacatt gagacagcat agaataagtg
     1981 cgacatcatc atcggaagag agtagtaaca aaggtcaaag acagttgact gtatcgccgg
     2041 aattcccggg gatcctatgt cttcagaaga agaagaagaa gaagcaaccc caggaggaaa
     2101 ggcagccact cctcatggac aaagacgact accacagctt gctgtatcag agccatctgt
     2161 gaacatccta ggaaacagag tgggactgaa aggttttacc tcactcgacc tatttgttgg
     2221 tgtttctaca aatttaaact agtataaaac atagaccata gctgtttggc tttttttcag
     2281 acccatgttt tttcctaagt cctagtttct aagaaatgac tgggatttgc taaaatatat
     2341 atatatataa ataataactt actaatagct aaataaaatt tcctcttaca actaattgag
     2401 ctggttttta tgaatgtgtc ttaattattt aaacttgagg cacatttttg ttttccttac
     2461 ttcattgtga atttccggaa ttctgcagcc aagctaattc cgggcgaatt tcttatgatt
     2521 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2581 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2641 gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgccggc
     2701 aagtgcacaa acaatactta aataaatact actcagtaat aacctatttc ttagcatttt
     2761 tgacgaaatt tgctattttg ttagagtctt ttacaccatt tgtctccaca cctccgctta
     2821 catcaacacc aataacgcca tttaatctaa gcgcatcacc aacattttct ggcgtcagtc
     2881 caccagctaa cataaaatgt aagctttcgg ggctctcttg ccttccaacc cagtcagaaa
     2941 tcgagttcca atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac aagggaataa
     3001 acgaatgagg tttctgtgaa gctgcactga gtagtatgtt gcagtctttt ggaaatacga
     3061 gtcttttaat aactggcaaa ccgaggaact cttggtattc ttgccacgac tcatctccat
     3121 gcagttggac gatatcaatg ccgtaatcat tgaccagagc caaaacatcc tccttaggtt
     3181 gattacgaaa cacgccaacc aagtatttcg gagtgcctga actattttta tatgctttta
     3241 caagacttga aattttcctt gcaataaccg ggtcaattgt tctctttcta ttgggcacac
     3301 atataatacc cagcaagtca gcatcggaat ctagagcaca ttctgcggcc tctgtgctct
     3361 gcaagccgca aactttcacc aatggaccag aactacctgt gaaattaata acagacatac
     3421 tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt caccaatgcc
     3481 ctccctcttg gccctctcct tttctttttt cgaccgaatt aattcgtaat catgtcatag
     3541 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3601 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3661 tcactgcccg ctttccagtc gggaaacctg tcgtgccagg aagatccgag gcctagcttc
     3721 taattcttcc aacatacaat gggagtttgg ccgagtggtt taaggcgtca gatttaggtg
     3781 gatttaacct ctaaaatctc tgatatcttc ggatgcaagg gttcgaatcc cttagctctc
     3841 attatttttt gctttttctc ttgaggtcac atgatcgcaa aatggcaaat ggcacgtgaa
     3901 gctgtcgata ttggggaact gtggtggttg gcaaatgact aattaagtta gtcaaggcgc
     3961 catcctcatg aaaactgtgt aacataataa ccgaagtgtc gaaaaggtgg caccttgtcc
     4021 aattgaacac gctcgatgaa aaaaataaga tatatataag gttaagtaaa gcgtctgtta
     4081 gaaaggaagt ttttcctttt tcttgctctc ttgtcttttc atctactatt tccttcgtgt
     4141 aatacagggt cgtcagatac atagatacaa ttctattacc cccatccata caatgggcca
     4201 tatggcttct agctatcctt atgacgtgcc tgactatgcc agcctgggag gaccttctag
     4261 tcctaagaag aagagaaagg tggcggccgc catgatccgc tttatcctca tccagaaccg
     4321 ggcaggcaag acacgcctgg ccaagtggta tatgcagttc gatgatgacg agaagcagaa
     4381 gttgatcgag gaagtgcacg ccgtggtcac cgtcggggat gccaagcaca ccaactttgt
     4441 ggagttccgg aacttcaaga tcatctaccg gcgctacgct ggcctctact tctgcatctg
     4501 cgttgacgtc aacgacaaca atctggccta tcttgaggcc atccacaact tcgtagaagt
     4561 gttaaatgaa tacttccaca atgtctgtga actggacctg gtgttcaact tctacaaggt
     4621 ttacacggtg gtagatgaga tgttcctggc aggagagatc cgagagacga gccagacgaa
     4681 ggtgctgaag cagctgctga tgctacagtc cctggagtga agatcttcgg gctgatctcc
     4741 catgtctcta ctggtggtgg tgcttctttg gaattattgg aaggtaagga attgccaggt
     4801 gttgctttct tatccgaaaa gaaataaatt gaattgaatt gaaatcgata gatcaatttt
     4861 tttcttttct ctttccccat cctttacgct aaaataatag tttattttat tttttgaata
     4921 ttttttattt atatacgtat atatagacta ttatttatct tttaatgatt attaagattt
     4981 ttattaaaaa aaaattcgct cctcttttaa tgcctttatg cagttttttt ttcccattcg
     5041 atatttctat gttcgggttc agcgtatttt aagtttaata actcgaaaat tctgcgttcg
     5101 ttaaagctag gcctcggatc ttcctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5161 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5221 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5281 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5341 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5401 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5461 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5521 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5581 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5641 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5701 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5761 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5821 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5881 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6121 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6181 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6241 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6301 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6361 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6421 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6481 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6541 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6601 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6661 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6721 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6781 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6841 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6901 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6961 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7021 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7081 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     7141 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     7201 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     7261 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     7321 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataacg
//
