LOCUS       sequence-258364-        7288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pTDH3_FdeR-N_2NLS_tPGK1-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  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 7288)
  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_004_026_096
FEATURES             Location/Qualifiers
     source          1..7288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        594..1260
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1130..1153
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2203..2223
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2227..2247
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4116..4919
                     /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(5561..6376)
                     /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(5561..6376)
                     /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     6284..6303
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6368..6387
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6601..7189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7090..7109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcca atgctaaatt cgagtgaaac acaggaagat cagaaaatcc tcatttcatc
       61 catattaaca ataatttcaa atgtttattt gcattatttg aaactaggca agacaagcaa
      121 cgaaacgttt ttgaaaattt tgagtatttt caataaattt gtagaggact cagatattga
      181 aaaaaagcta cagcaattaa tacttgataa gaagagtatt gagaagggca acggttcatc
      241 atctcatgga tctgcacatg aacaaacacc agagtcaaac gacgttgaaa ttgaggctac
      301 tgcgccaatt gatgacaata cagacgatga taacaaaccg aagttatctg atgtagaaaa
      361 ggattaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt ctatatatgt
      421 taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc atcaaagaag
      481 gttaatgtgg ctgtggtttc agggtccata aagcccacat ggataacatt acccctgaat
      541 tcgcatctag actgaactgg ccgataattg cagacgaacg cagttcgagt ttatcattat
      601 caatactgcc atttcaaaga atacgtaaat aattaatagt agtgattttc ctaactttat
      661 ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc
      721 gggttacaca gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc
      781 actaaatata atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca
      841 ccaaaatatt gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta
      901 cagagaacag gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc
      961 tgcctggagt aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct
     1021 tacaccttct attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa
     1081 accagttccc tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt
     1141 gattgtaatt ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct
     1201 tttttttagt tttaaaacac caagaactta gtttcgaata aacacacata aacaaacaaa
     1261 agatctatgc gtttcaacaa gctcgacctc aatcttctgg tcgccctgga tgcactgctc
     1321 acggagatga gcatcagccg cgccgccgaa aagatccatc tgagccagtc ggccatgagc
     1381 aatgccctgg cgcggctgcg cgagtatttc gatgatgaat tgctgatcca ggtgggccgg
     1441 cgcatggagc ccacgccgcg cgccgaggtg ctcaaggatg cggtgcatga tgtgctgcgg
     1501 cgtatcgatg gctccatcgc ggcgctgccg gccttcgtgc cggccgagtc cacgcgcgag
     1561 tttcgcatct cggtttcgga ctttacgctc tccgtcctca tcccccgggt gctggcgcgc
     1621 gcgcacgccg agggcaagca catccgcttt gccctgatgc cgcaggtgca agacccgacc
     1681 cgctcgctgg atcgggccga ggtggacctg ctggtcttgc cgcaggaatt ctgcacgccc
     1741 gatcatcctg ccgaagaggt cttccgcgaa cggcatgtct gcgtggtctg gcgcgacagt
     1801 gcgctggcgc aaggcgagct gacgctggaa cgctacatgg cctcaggcca tgtggtgatg
     1861 gtgccgcctg gggccaatgc gtcgtcggtg gaggcgtgga tggccaggaa gctgggcttt
     1921 gcgcgccggg tggaagtgac cagcttcagc ttcgcttctg cgctggcgct ggtacagggg
     1981 acggaccgca tcgccacggt gcatgcccgg ctggcgcagc tgctggctcc gcaatggccg
     2041 gtggtgatca aggagagtcc gctgtcgctg ggcgagatgc ggcagatgat gcagtggcat
     2101 cgctaccgca gcaatgatcc tggcatccag tggctgcgtc gggtgtttct ggagagtgcg
     2161 caggagatgg atgcggcgct gccaggcatc tgcggatccg atccaaaaaa gaagagaaag
     2221 gtagatccaa aaaagaagag aaaggtataa ctcgagtggc attgaattga attgaaatcg
     2281 atagatcaat ttttttcttt tctctttccc catcctttac gctaaaataa tagtttattt
     2341 tattttttga atatttttta tttatatacg tatatataga ctattattta tcttttaatg
     2401 attattaaga tttttattaa aaaaaaattc gctcctcttt taatgccttt atgcagtttt
     2461 tttttcccat tcgatatttc tatgtgctgc caaaccagat gtcaacacag ctacaacgaa
     2521 gcttcgtctt ttcttgaaga aaagaggaag ctcgctaaat gggattccac tttccgttcc
     2581 ctgccagctg atggaaaaag gttagtggaa cgatgaagaa taaaaagaga gatccactga
     2641 ggtgaaattt cagctgacag cgagtttcat gatcgtgatg aacaatggta acgagttgtg
     2701 gctgttgcca gggagggtgg ttctcaccaa atcgctactt gggtttgaga ttggtaaaat
     2761 tgattgtttg gatggcaatc atctatggaa tggatattat ataacaaaga agaaataatg
     2821 aactgattct cttcctcctt cttgtccttt cttaattctg ttgtaattac cttcctttgt
     2881 aatttttttt gtaattattc ttcttaataa tccaaacaaa cacacatatt acaataagat
     2941 ctatggtttc aaagggcgaa gaagacaata tggctattat taaggaattc atgagattca
     3001 aggtccacat ggaaggttct gtcaacggtc acgaattcga aattgaaggt gaaggtgaag
     3061 gtagaccata cgaaggtacc caaaccgcta agttgaaggt caccaagggt ggtccattgc
     3121 cattcgcttg ggatattttg tctccacaat tcatgtacgg ttctaaggct tacgtcaagc
     3181 acccagctga tattccagat tacttgaagt tgtctttccc agaaggtttc aagtgggaaa
     3241 gagtcatgaa cttcgaagat ggtggtgtcg tcaccgtcac ccaagattct tctttgcaag
     3301 atggtgaatt catttacaag gtcaagttga gaggtaccaa cttcccatct gatggtccag
     3361 tcatgcaaaa gaagaccatg ggttgggaag cttcttctga aagaatgtac ccagaagatg
     3421 gtgctttgaa gggtgaaatt aagcaaagat tgaagttgaa ggatggtggt cactacgatg
     3481 ctgaagtcaa gaccacctac aaggctaaga agccagtcca attgccaggt gcttacaacg
     3541 tcaacattaa gttggatatt acctctcaca acgaagatta caccattgtc gaacaatacg
     3601 aaagagctga aggtagacac tctaccggtg gtatggatga attgtacaag taaatcctaa
     3661 ctcgagataa agcaatcttg atgaggataa tgattttttt ttgaatatac ataaatacta
     3721 ccgtttttct gctagatttt gtgatgacgt aaataagtac atattacttt ttaagccaag
     3781 acaagattaa gcattaactt tacccttttc tttctaagtt tcaatattag ttatcactgt
     3841 ttaaaagtta tggcgagaac gtcggcggtt aaaatatatt accctgaacg gctgagcagt
     3901 tacagagatg ttacgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
     3961 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     4021 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     4081 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     4141 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     4201 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     4261 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     4321 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     4381 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     4441 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     4501 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     4561 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     4621 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     4681 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     4741 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     4801 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     4861 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     4921 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     4981 tatatcagtt attacccgag tagagcactt gaatccactg ccccgggaat ctcggtcgta
     5041 atgatttcta taatgacgaa aaaaaaaaaa ttggaaagaa aaagcttcat ggcctttata
     5101 aaaaggaact atccaatacc tcgccagaac caagtaacag tattttacgg ggcacaaatc
     5161 aagaacaata agacaggact gtaaagatgg acgcattgaa ctccaaagaa caacaagagt
     5221 tccaaaaagt agtggaacaa aagcaaatga aggatttcat gcgtttgtac tctaatctgg
     5281 tagaaagatg tttcacagac tgtgtcaatg acttcacaac atcaaagcta accaataagg
     5341 aacaaacatg catcatgaag tgctcagaaa agttcttgaa gcatagcgaa cgtgtagggc
     5401 agcgtttcca agaacaaaac gctgccttgg gacaaggctt gggccgataa ggtgtactgg
     5461 cgtatatata tctaattatg tatctctggt gtagcccatt tttagcatgt aaatataaag
     5521 accgagcggc cgcgtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     5581 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     5641 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     5701 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     5761 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     5821 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     5881 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg aggcgaaata
     5941 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     6001 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     6061 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     6121 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     6181 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     6241 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     6301 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc
     6361 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     6421 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     6481 ggctttgttg aataaatcga acttttgctg agttgaagga tcagtcatga ccaaaatccc
     6541 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6601 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6661 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6721 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6781 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6841 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6901 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6961 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7021 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7081 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7141 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7201 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7261 gttatcccct gattctgtgg ataaccgt
//
