LOCUS       sequence-258361-        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        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             1270..1290
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4092..4895
                     /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(5537..6352)
                     /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(5537..6352)
                     /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     6260..6279
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6344..6363
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6577..7165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7066..7085
                     /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 caaagaagaa gagaaaggtt ggttctcgtt tcaacaagct cgacctcaat
     1321 cttctggtcg ccctggatgc actgctcacg gagatgagca tcagccgcgc cgccgaaaag
     1381 atccatctga gccagtcggc catgagcaat gccctggcgc ggctgcgcga gtatttcgat
     1441 gatgaattgc tgatccaggt gggccggcgc atggagccca cgccgcgcgc cgaggtgctc
     1501 aaggatgcgg tgcatgatgt gctgcggcgt atcgatggct ccatcgcggc gctgccggcc
     1561 ttcgtgccgg ccgagtccac gcgcgagttt cgcatctcgg tttcggactt tacgctctcc
     1621 gtcctcatcc cccgggtgct ggcgcgcgcg cacgccgagg gcaagcacat ccgctttgcc
     1681 ctgatgccgc aggtgcaaga cccgacccgc tcgctggatc gggccgaggt ggacctgctg
     1741 gtcttgccgc aggaattctg cacgcccgat catcctgccg aagaggtctt ccgcgaacgg
     1801 catgtctgcg tggtctggcg cgacagtgcg ctggcgcaag gcgagctgac gctggaacgc
     1861 tacatggcct caggccatgt ggtgatggtg ccgcctgggg ccaatgcgtc gtcggtggag
     1921 gcgtggatgg ccaggaagct gggctttgcg cgccgggtgg aagtgaccag cttcagcttc
     1981 gcttctgcgc tggcgctggt acaggggacg gaccgcatcg ccacggtgca tgcccggctg
     2041 gcgcagctgc tggctccgca atggccggtg gtgatcaagg agagtccgct gtcgctgggc
     2101 gagatgcggc agatgatgca gtggcatcgc taccgcagca atgatcctgg catccagtgg
     2161 ctgcgtcggg tgtttctgga gagtgcgcag gagatggatg cggcgctgcc aggcatctgc
     2221 tgaatcctaa ctcgagattg aattgaattg aaatcgatag atcaattttt ttcttttctc
     2281 tttccccatc ctttacgcta aaataatagt ttattttatt ttttgaatat tttttattta
     2341 tatacgtata tatagactat tatttatctt ttaatgatta ttaagatttt tattaaaaaa
     2401 aaattcgctc ctcttttaat gcctttatgc agtttttttt tcccattcga tatttctatg
     2461 tgctgccaaa ccagatgtca acacagctac aacgaagctt cgtcttttct tgaagaaaag
     2521 aggaagctcg ctaaatggga ttccactttc cgttccctgc cagctgatgg aaaaaggtta
     2581 gtggaacgat gaagaataaa aagagagatc cactgaggtg aaatttcagc tgacagcgag
     2641 tttcatgatc gtgatgaaca atggtaacga gttgtggctg ttgccaggga gggtggttct
     2701 caccaaatcg ctacttgggt ttgagattgg taaaattgat tgtttggatg gcaatcatct
     2761 atggaatgga tattatataa caaagaagaa ataatgaact gattctcttc ctccttcttg
     2821 tcctttctta attctgttgt aattaccttc ctttgtaatt ttttttgtaa ttattcttct
     2881 taataatcca aacaaacaca catattacaa taagatctat ggtttcaaag ggcgaagaag
     2941 acaatatggc tattattaag gaattcatga gattcaaggt ccacatggaa ggttctgtca
     3001 acggtcacga attcgaaatt gaaggtgaag gtgaaggtag accatacgaa ggtacccaaa
     3061 ccgctaagtt gaaggtcacc aagggtggtc cattgccatt cgcttgggat attttgtctc
     3121 cacaattcat gtacggttct aaggcttacg tcaagcaccc agctgatatt ccagattact
     3181 tgaagttgtc tttcccagaa ggtttcaagt gggaaagagt catgaacttc gaagatggtg
     3241 gtgtcgtcac cgtcacccaa gattcttctt tgcaagatgg tgaattcatt tacaaggtca
     3301 agttgagagg taccaacttc ccatctgatg gtccagtcat gcaaaagaag accatgggtt
     3361 gggaagcttc ttctgaaaga atgtacccag aagatggtgc tttgaagggt gaaattaagc
     3421 aaagattgaa gttgaaggat ggtggtcact acgatgctga agtcaagacc acctacaagg
     3481 ctaagaagcc agtccaattg ccaggtgctt acaacgtcaa cattaagttg gatattacct
     3541 ctcacaacga agattacacc attgtcgaac aatacgaaag agctgaaggt agacactcta
     3601 ccggtggtat ggatgaattg tacaagtaaa tcctaactcg agataaagca atcttgatga
     3661 ggataatgat ttttttttga atatacataa atactaccgt ttttctgcta gattttgtga
     3721 tgacgtaaat aagtacatat tactttttaa gccaagacaa gattaagcat taactttacc
     3781 cttttctttc taagtttcaa tattagttat cactgtttaa aagttatggc gagaacgtcg
     3841 gcggttaaaa tatattaccc tgaacggctg agcagttaca gagatgttac gaaccactag
     3901 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     3961 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     4021 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     4081 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     4141 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     4201 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     4261 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     4321 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     4381 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     4441 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     4501 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     4561 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     4621 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     4681 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     4741 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     4801 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     4861 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     4921 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaga
     4981 gcacttgaat ccactgcccc gggaatctcg gtcgtaatga tttctataat gacgaaaaaa
     5041 aaaaaattgg aaagaaaaag cttcatggcc tttataaaaa ggaactatcc aatacctcgc
     5101 cagaaccaag taacagtatt ttacggggca caaatcaaga acaataagac aggactgtaa
     5161 agatggacgc attgaactcc aaagaacaac aagagttcca aaaagtagtg gaacaaaagc
     5221 aaatgaagga tttcatgcgt ttgtactcta atctggtaga aagatgtttc acagactgtg
     5281 tcaatgactt cacaacatca aagctaacca ataaggaaca aacatgcatc atgaagtgct
     5341 cagaaaagtt cttgaagcat agcgaacgtg tagggcagcg tttccaagaa caaaacgctg
     5401 ccttgggaca aggcttgggc cgataaggtg tactggcgta tatatatcta attatgtatc
     5461 tctggtgtag cccattttta gcatgtaaat ataaagaccg agcggccgcg tgttacaacc
     5521 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     5581 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     5641 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     5701 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     5761 caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga
     5821 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     5881 tattcattcg tgattgcgcc tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat
     5941 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     6001 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     6061 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     6121 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     6181 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     6241 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     6301 tggaatttaa tcgcggcctg gagcaagacg tttcccgttg aatatggctc ataacacccc
     6361 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     6421 gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt
     6481 ttgctgagtt gaaggatcag tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6541 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6601 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6661 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6721 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6781 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6841 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6901 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6961 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7021 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7081 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7141 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7201 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7261 ccgt
//
