LOCUS       sequence-258368-        7315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pTDH3_ NLS_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 7315)
  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 7315)
  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_007_026_096
FEATURES             Location/Qualifiers
     source          1..7315
                     /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             2230..2250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2254..2274
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4143..4946
                     /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(5588..6403)
                     /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(5588..6403)
                     /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     6311..6330
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6395..6414
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6628..7216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7117..7136
                     /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 ggatccgatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtataactc
     2281 gagtggcatt gaattgaatt gaaatcgata gatcaatttt tttcttttct ctttccccat
     2341 cctttacgct aaaataatag tttattttat tttttgaata ttttttattt atatacgtat
     2401 atatagacta ttatttatct tttaatgatt attaagattt ttattaaaaa aaaattcgct
     2461 cctcttttaa tgcctttatg cagttttttt ttcccattcg atatttctat gtgctgccaa
     2521 accagatgtc aacacagcta caacgaagct tcgtcttttc ttgaagaaaa gaggaagctc
     2581 gctaaatggg attccacttt ccgttccctg ccagctgatg gaaaaaggtt agtggaacga
     2641 tgaagaataa aaagagagat ccactgaggt gaaatttcag ctgacagcga gtttcatgat
     2701 cgtgatgaac aatggtaacg agttgtggct gttgccaggg agggtggttc tcaccaaatc
     2761 gctacttggg tttgagattg gtaaaattga ttgtttggat ggcaatcatc tatggaatgg
     2821 atattatata acaaagaaga aataatgaac tgattctctt cctccttctt gtcctttctt
     2881 aattctgttg taattacctt cctttgtaat tttttttgta attattcttc ttaataatcc
     2941 aaacaaacac acatattaca ataagatcta tggtttcaaa gggcgaagaa gacaatatgg
     3001 ctattattaa ggaattcatg agattcaagg tccacatgga aggttctgtc aacggtcacg
     3061 aattcgaaat tgaaggtgaa ggtgaaggta gaccatacga aggtacccaa accgctaagt
     3121 tgaaggtcac caagggtggt ccattgccat tcgcttggga tattttgtct ccacaattca
     3181 tgtacggttc taaggcttac gtcaagcacc cagctgatat tccagattac ttgaagttgt
     3241 ctttcccaga aggtttcaag tgggaaagag tcatgaactt cgaagatggt ggtgtcgtca
     3301 ccgtcaccca agattcttct ttgcaagatg gtgaattcat ttacaaggtc aagttgagag
     3361 gtaccaactt cccatctgat ggtccagtca tgcaaaagaa gaccatgggt tgggaagctt
     3421 cttctgaaag aatgtaccca gaagatggtg ctttgaaggg tgaaattaag caaagattga
     3481 agttgaagga tggtggtcac tacgatgctg aagtcaagac cacctacaag gctaagaagc
     3541 cagtccaatt gccaggtgct tacaacgtca acattaagtt ggatattacc tctcacaacg
     3601 aagattacac cattgtcgaa caatacgaaa gagctgaagg tagacactct accggtggta
     3661 tggatgaatt gtacaagtaa atcctaactc gagataaagc aatcttgatg aggataatga
     3721 tttttttttg aatatacata aatactaccg tttttctgct agattttgtg atgacgtaaa
     3781 taagtacata ttacttttta agccaagaca agattaagca ttaactttac ccttttcttt
     3841 ctaagtttca atattagtta tcactgttta aaagttatgg cgagaacgtc ggcggttaaa
     3901 atatattacc ctgaacggct gagcagttac agagatgtta cgaaccacta gtgcactgca
     3961 gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     4021 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     4081 attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa
     4141 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     4201 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     4261 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     4321 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     4381 tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata
     4441 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     4501 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     4561 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     4621 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     4681 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     4741 tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg
     4801 tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     4861 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     4921 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     4981 actcacaaat tagagcttca atttaattat atcagttatt acccgagtag agcacttgaa
     5041 tccactgccc cgggaatctc ggtcgtaatg atttctataa tgacgaaaaa aaaaaaattg
     5101 gaaagaaaaa gcttcatggc ctttataaaa aggaactatc caatacctcg ccagaaccaa
     5161 gtaacagtat tttacggggc acaaatcaag aacaataaga caggactgta aagatggacg
     5221 cattgaactc caaagaacaa caagagttcc aaaaagtagt ggaacaaaag caaatgaagg
     5281 atttcatgcg tttgtactct aatctggtag aaagatgttt cacagactgt gtcaatgact
     5341 tcacaacatc aaagctaacc aataaggaac aaacatgcat catgaagtgc tcagaaaagt
     5401 tcttgaagca tagcgaacgt gtagggcagc gtttccaaga acaaaacgct gccttgggac
     5461 aaggcttggg ccgataaggt gtactggcgt atatatatct aattatgtat ctctggtgta
     5521 gcccattttt agcatgtaaa tataaagacc gagcggccgc gtgttacaac caattaacca
     5581 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     5641 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     5701 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     5761 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     5821 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     5881 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     5941 gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     6001 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     6061 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     6121 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     6181 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     6241 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     6301 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     6361 atcgcggcct ggagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     6421 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     6481 aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt
     6541 tgaaggatca gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7261 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgt
//
