LOCUS       sequence-267008-        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        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             1660..1680
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1684..1704
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3573..4376
                     /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(5018..5833)
                     /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(5018..5833)
                     /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     5741..5760
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5825..5844
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6058..6646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6547..6566
                     /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 tctatgcgtt tcaacaagct cgacctcaat cttctggtcg ccctggatgc actgctcacg
      781 gagatgagca tcagccgcgc cgccgaaaag atccatctga gccagtcggc catgagcaat
      841 gccctggcgc ggctgcgcga gtatttcgat gatgaattgc tgatccaggt gggccggcgc
      901 atggagccca cgccgcgcgc cgaggtgctc aaggatgcgg tgcatgatgt gctgcggcgt
      961 atcgatggct ccatcgcggc gctgccggcc ttcgtgccgg ccgagtccac gcgcgagttt
     1021 cgcatctcgg tttcggactt tacgctctcc gtcctcatcc cccgggtgct ggcgcgcgcg
     1081 cacgccgagg gcaagcacat ccgctttgcc ctgatgccgc aggtgcaaga cccgacccgc
     1141 tcgctggatc gggccgaggt ggacctgctg gtcttgccgc aggaattctg cacgcccgat
     1201 catcctgccg aagaggtctt ccgcgaacgg catgtctgcg tggtctggcg cgacagtgcg
     1261 ctggcgcaag gcgagctgac gctggaacgc tacatggcct caggccatgt ggtgatggtg
     1321 ccgcctgggg ccaatgcgtc gtcggtggag gcgtggatgg ccaggaagct gggctttgcg
     1381 cgccgggtgg aagtgaccag cttcagcttc gcttctgcgc tggcgctggt acaggggacg
     1441 gaccgcatcg ccacggtgca tgcccggctg gcgcagctgc tggctccgca atggccggtg
     1501 gtgatcaagg agagtccgct gtcgctgggc gagatgcggc agatgatgca gtggcatcgc
     1561 taccgcagca atgatcctgg catccagtgg ctgcgtcggg tgtttctgga gagtgcgcag
     1621 gagatggatg cggcgctgcc aggcatctgc ggatccgatc caaaaaagaa gagaaaggta
     1681 gatccaaaaa agaagagaaa ggtataactc gagtggcatt gaattgaatt gaaatcgata
     1741 gatcaatttt tttcttttct ctttccccat cctttacgct aaaataatag tttattttat
     1801 tttttgaata ttttttattt atatacgtat atatagacta ttatttatct tttaatgatt
     1861 attaagattt ttattaaaaa aaaattcgct cctcttttaa tgcctttatg cagttttttt
     1921 ttcccattcg atatttctat gtgctgccaa accagatgtc aacacagcta caacgaagct
     1981 tcgtcttttc ttgaagaaaa gaggaagctc gctaaatggg attccacttt ccgttccctg
     2041 ccagctgatg gaaaaaggtt agtggaacga tgaagaataa aaagagagat ccactgaggt
     2101 gaaatttcag ctgacagcga gtttcatgat cgtgatgaac aatggtaacg agttgtggct
     2161 gttgccaggg agggtggttc tcaccaaatc gctacttggg tttgagattg gtaaaattga
     2221 ttgtttggat ggcaatcatc tatggaatgg atattatata acaaagaaga aataatgaac
     2281 tgattctctt cctccttctt gtcctttctt aattctgttg taattacctt cctttgtaat
     2341 tttttttgta attattcttc ttaataatcc aaacaaacac acatattaca ataagatcta
     2401 tggtttcaaa gggcgaagaa gacaatatgg ctattattaa ggaattcatg agattcaagg
     2461 tccacatgga aggttctgtc aacggtcacg aattcgaaat tgaaggtgaa ggtgaaggta
     2521 gaccatacga aggtacccaa accgctaagt tgaaggtcac caagggtggt ccattgccat
     2581 tcgcttggga tattttgtct ccacaattca tgtacggttc taaggcttac gtcaagcacc
     2641 cagctgatat tccagattac ttgaagttgt ctttcccaga aggtttcaag tgggaaagag
     2701 tcatgaactt cgaagatggt ggtgtcgtca ccgtcaccca agattcttct ttgcaagatg
     2761 gtgaattcat ttacaaggtc aagttgagag gtaccaactt cccatctgat ggtccagtca
     2821 tgcaaaagaa gaccatgggt tgggaagctt cttctgaaag aatgtaccca gaagatggtg
     2881 ctttgaaggg tgaaattaag caaagattga agttgaagga tggtggtcac tacgatgctg
     2941 aagtcaagac cacctacaag gctaagaagc cagtccaatt gccaggtgct tacaacgtca
     3001 acattaagtt ggatattacc tctcacaacg aagattacac cattgtcgaa caatacgaaa
     3061 gagctgaagg tagacactct accggtggta tggatgaatt gtacaagtaa atcctaactc
     3121 gagataaagc aatcttgatg aggataatga tttttttttg aatatacata aatactaccg
     3181 tttttctgct agattttgtg atgacgtaaa taagtacata ttacttttta agccaagaca
     3241 agattaagca ttaactttac ccttttcttt ctaagtttca atattagtta tcactgttta
     3301 aaagttatgg cgagaacgtc ggcggttaaa atatattacc ctgaacggct gagcagttac
     3361 agagatgtta cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     3421 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     3481 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     3541 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     3601 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     3661 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     3721 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     3781 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     3841 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     3901 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     3961 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     4021 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     4081 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     4141 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     4201 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     4261 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     4321 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     4381 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     4441 atcagttatt acccgagtag agcacttgaa tccactgccc cgggaatctc ggtcgtaatg
     4501 atttctataa tgacgaaaaa aaaaaaattg gaaagaaaaa gcttcatggc ctttataaaa
     4561 aggaactatc caatacctcg ccagaaccaa gtaacagtat tttacggggc acaaatcaag
     4621 aacaataaga caggactgta aagatggacg cattgaactc caaagaacaa caagagttcc
     4681 aaaaagtagt ggaacaaaag caaatgaagg atttcatgcg tttgtactct aatctggtag
     4741 aaagatgttt cacagactgt gtcaatgact tcacaacatc aaagctaacc aataaggaac
     4801 aaacatgcat catgaagtgc tcagaaaagt tcttgaagca tagcgaacgt gtagggcagc
     4861 gtttccaaga acaaaacgct gccttgggac aaggcttggg ccgataaggt gtactggcgt
     4921 atatatatct aattatgtat ctctggtgta gcccattttt agcatgtaaa tataaagacc
     4981 gagcggccgc gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca
     5041 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     5101 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     5161 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     5221 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     5281 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     5341 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc
     5401 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     5461 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     5521 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     5581 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     5641 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     5701 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     5761 catataaatc agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt
     5821 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     5881 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc
     5941 tttgttgaat aaatcgaact tttgctgagt tgaaggatca gtcatgacca aaatccctta
     6001 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6061 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6121 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6181 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6241 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6301 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6361 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6421 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6481 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6541 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6601 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6661 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6721 atcccctgat tctgtggata accgtgcggc cgccaatgct aaattcgagt gaaacacagg
     6781 aagatcagaa aatcctcatt tcatccatat taacaataat ttcaaatgtt tatttgcatt
     6841 atttgaaact aggcaagaca agcaacgaaa cgtttttgaa aattttgagt attttcaata
     6901 aatttgtaga ggactcagat attgaaaaaa agctacagca attaatactt gataagaaga
     6961 gtattgagaa gggcaacggt tcatcatctc atggatctgc acatgaacaa acaccagagt
     7021 caaacgacgt tgaaattgag gctactgcgc caattgatga caatacagac gatgataaca
     7081 aaccgaagtt atctgatgta gaaaaggatt aaagatgcta agagatagtg atgatatttc
     7141 ataaataatg taattctata tatgttaatt accttttttg cgaggcatat ttatggtgaa
     7201 ggataagttt tgaccatcaa agaaggttaa tgtggctgtg gtttcagggt ccataaagcc
     7261 cacatggata acattacccc tgaattcg
//
