LOCUS       sequence-267010-        7264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pTDH3_NLS_FdeR-N_tPGK1-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7264)
  AUTHORS   Wang R, Cress BF, Yang Z, Hordines JC 3rd, Zhao S, Jung GY, Wang Z, 
            Koffas MAG
  TITLE     Design and Characterization of Biosensors for the Screening of 
            Modular Assembled Naringenin Biosynthetic Library in Saccharomyces 
            cerevisiae.
  JOURNAL   ACS Synth Biol. 2019 Sep 20;8(9):2121-2130. doi: 
            10.1021/acssynbio.9b00212. Epub 2019 Aug 28.
  PUBMED    31433622
REFERENCE   2  (bases 1 to 7264)
  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: 
            "10.1021/acssynbio.9b00212"; journalName: "ACS Synth Biol"; date: 
            "2019-09-20- 20"; volume: "8"; issue: "9"; pages: "2121-2130"
COMMENT     Sequence Label: S3_001_026_096
FEATURES             Location/Qualifiers
     source          1..7264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        51..717
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     587..610
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             727..747
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3549..4352
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(4994..5809)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4994..5809)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5717..5736
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5801..5820
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6034..6622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6523..6542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catctagact gaactggccg ataattgcag acgaacgcag ttcgagttta tcattatcaa
       61 tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta actttattta
      121 gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg
      181 ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact
      241 aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca
      301 aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag
      361 agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc
      421 ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac
      481 accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc
      541 agttccctga aattattccc ctacttgact aataagtata taaagacggt aggtattgat
      601 tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt
      661 ttttagtttt aaaacaccaa gaacttagtt tcgaataaac acacataaac aaacaaaaga
      721 tctatgccaa agaagaagag aaaggttggt tctcgtttca acaagctcga cctcaatctt
      781 ctggtcgccc tggatgcact gctcacggag atgagcatca gccgcgccgc cgaaaagatc
      841 catctgagcc agtcggccat gagcaatgcc ctggcgcggc tgcgcgagta tttcgatgat
      901 gaattgctga tccaggtggg ccggcgcatg gagcccacgc cgcgcgccga ggtgctcaag
      961 gatgcggtgc atgatgtgct gcggcgtatc gatggctcca tcgcggcgct gccggccttc
     1021 gtgccggccg agtccacgcg cgagtttcgc atctcggttt cggactttac gctctccgtc
     1081 ctcatccccc gggtgctggc gcgcgcgcac gccgagggca agcacatccg ctttgccctg
     1141 atgccgcagg tgcaagaccc gacccgctcg ctggatcggg ccgaggtgga cctgctggtc
     1201 ttgccgcagg aattctgcac gcccgatcat cctgccgaag aggtcttccg cgaacggcat
     1261 gtctgcgtgg tctggcgcga cagtgcgctg gcgcaaggcg agctgacgct ggaacgctac
     1321 atggcctcag gccatgtggt gatggtgccg cctggggcca atgcgtcgtc ggtggaggcg
     1381 tggatggcca ggaagctggg ctttgcgcgc cgggtggaag tgaccagctt cagcttcgct
     1441 tctgcgctgg cgctggtaca ggggacggac cgcatcgcca cggtgcatgc ccggctggcg
     1501 cagctgctgg ctccgcaatg gccggtggtg atcaaggaga gtccgctgtc gctgggcgag
     1561 atgcggcaga tgatgcagtg gcatcgctac cgcagcaatg atcctggcat ccagtggctg
     1621 cgtcgggtgt ttctggagag tgcgcaggag atggatgcgg cgctgccagg catctgctga
     1681 atcctaactc gagattgaat tgaattgaaa tcgatagatc aatttttttc ttttctcttt
     1741 ccccatcctt tacgctaaaa taatagttta ttttattttt tgaatatttt ttatttatat
     1801 acgtatatat agactattat ttatctttta atgattatta agatttttat taaaaaaaaa
     1861 ttcgctcctc ttttaatgcc tttatgcagt ttttttttcc cattcgatat ttctatgtgc
     1921 tgccaaacca gatgtcaaca cagctacaac gaagcttcgt cttttcttga agaaaagagg
     1981 aagctcgcta aatgggattc cactttccgt tccctgccag ctgatggaaa aaggttagtg
     2041 gaacgatgaa gaataaaaag agagatccac tgaggtgaaa tttcagctga cagcgagttt
     2101 catgatcgtg atgaacaatg gtaacgagtt gtggctgttg ccagggaggg tggttctcac
     2161 caaatcgcta cttgggtttg agattggtaa aattgattgt ttggatggca atcatctatg
     2221 gaatggatat tatataacaa agaagaaata atgaactgat tctcttcctc cttcttgtcc
     2281 tttcttaatt ctgttgtaat taccttcctt tgtaattttt tttgtaatta ttcttcttaa
     2341 taatccaaac aaacacacat attacaataa gatctatggt ttcaaagggc gaagaagaca
     2401 atatggctat tattaaggaa ttcatgagat tcaaggtcca catggaaggt tctgtcaacg
     2461 gtcacgaatt cgaaattgaa ggtgaaggtg aaggtagacc atacgaaggt acccaaaccg
     2521 ctaagttgaa ggtcaccaag ggtggtccat tgccattcgc ttgggatatt ttgtctccac
     2581 aattcatgta cggttctaag gcttacgtca agcacccagc tgatattcca gattacttga
     2641 agttgtcttt cccagaaggt ttcaagtggg aaagagtcat gaacttcgaa gatggtggtg
     2701 tcgtcaccgt cacccaagat tcttctttgc aagatggtga attcatttac aaggtcaagt
     2761 tgagaggtac caacttccca tctgatggtc cagtcatgca aaagaagacc atgggttggg
     2821 aagcttcttc tgaaagaatg tacccagaag atggtgcttt gaagggtgaa attaagcaaa
     2881 gattgaagtt gaaggatggt ggtcactacg atgctgaagt caagaccacc tacaaggcta
     2941 agaagccagt ccaattgcca ggtgcttaca acgtcaacat taagttggat attacctctc
     3001 acaacgaaga ttacaccatt gtcgaacaat acgaaagagc tgaaggtaga cactctaccg
     3061 gtggtatgga tgaattgtac aagtaaatcc taactcgaga taaagcaatc ttgatgagga
     3121 taatgatttt tttttgaata tacataaata ctaccgtttt tctgctagat tttgtgatga
     3181 cgtaaataag tacatattac tttttaagcc aagacaagat taagcattaa ctttaccctt
     3241 ttctttctaa gtttcaatat tagttatcac tgtttaaaag ttatggcgag aacgtcggcg
     3301 gttaaaatat attaccctga acggctgagc agttacagag atgttacgaa ccactagtgc
     3361 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
     3421 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     3481 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa
     3541 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     3601 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     3661 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     3721 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     3781 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg
     3841 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     3901 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     3961 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     4021 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     4081 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     4141 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata
     4201 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat
     4261 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     4321 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     4381 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtagagca
     4441 cttgaatcca ctgccccggg aatctcggtc gtaatgattt ctataatgac gaaaaaaaaa
     4501 aaattggaaa gaaaaagctt catggccttt ataaaaagga actatccaat acctcgccag
     4561 aaccaagtaa cagtatttta cggggcacaa atcaagaaca ataagacagg actgtaaaga
     4621 tggacgcatt gaactccaaa gaacaacaag agttccaaaa agtagtggaa caaaagcaaa
     4681 tgaaggattt catgcgtttg tactctaatc tggtagaaag atgtttcaca gactgtgtca
     4741 atgacttcac aacatcaaag ctaaccaata aggaacaaac atgcatcatg aagtgctcag
     4801 aaaagttctt gaagcatagc gaacgtgtag ggcagcgttt ccaagaacaa aacgctgcct
     4861 tgggacaagg cttgggccga taaggtgtac tggcgtatat atatctaatt atgtatctct
     4921 ggtgtagccc atttttagca tgtaaatata aagaccgagc ggccgcgtgt tacaaccaat
     4981 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     5041 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     5101 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     5161 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     5221 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     5281 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     5341 tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac
     5401 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     5461 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     5521 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     5581 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     5641 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     5701 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     5761 aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg
     5821 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     5881 caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg
     5941 ctgagttgaa ggatcagtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6721 tgcggccgcc aatgctaaat tcgagtgaaa cacaggaaga tcagaaaatc ctcatttcat
     6781 ccatattaac aataatttca aatgtttatt tgcattattt gaaactaggc aagacaagca
     6841 acgaaacgtt tttgaaaatt ttgagtattt tcaataaatt tgtagaggac tcagatattg
     6901 aaaaaaagct acagcaatta atacttgata agaagagtat tgagaagggc aacggttcat
     6961 catctcatgg atctgcacat gaacaaacac cagagtcaaa cgacgttgaa attgaggcta
     7021 ctgcgccaat tgatgacaat acagacgatg ataacaaacc gaagttatct gatgtagaaa
     7081 aggattaaag atgctaagag atagtgatga tatttcataa ataatgtaat tctatatatg
     7141 ttaattacct tttttgcgag gcatatttat ggtgaaggat aagttttgac catcaaagaa
     7201 ggttaatgtg gctgtggttt cagggtccat aaagcccaca tggataacat tacccctgaa
     7261 ttcg
//
