LOCUS       sequence-266907-        7245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV2-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 7245)
  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 7245)
  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_MV2
FEATURES             Location/Qualifiers
     source          1..7245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..667
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     537..560
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             1610..1630
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1634..1654
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3523..4326
                     /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"
     rep_origin      4409..5391
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5449..6264)
                     /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(5449..6264)
                     /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     6172..6191
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6256..6275
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6489..7077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6978..6997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcattatcaa tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta
       61 actttattta gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat
      121 agggggcggg ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct
      181 ggcatccact aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat
      241 cccagcacca aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc
      301 gcaactacag agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga
      361 tgcaacctgc ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc
      421 attttcttac accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa
      481 ggttgaaacc agttccctga aattattccc ctacttgact aataagtata taaagacggt
      541 aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag
      601 ttagtctttt ttttagtttt aaaacaccaa gaacttagtt tcgaataaac acacataaac
      661 aaacaaaaga tctatgcgtt tcaacaagct cgacctcaat cttctggtcg ccctggatgc
      721 actgctcacg gagatgagca tcagccgcgc cgccgaaaag atccatctga gccagtcggc
      781 catgagcaat gccctggcgc ggctgcgcga gtatttcgat gatgaattgc tgatccaggt
      841 gggccggcgc atggagccca cgccgcgcgc cgaggtgctc aaggatgcgg tgcatgatgt
      901 gctgcggcgt atcgatggct ccatcgcggc gctgccggcc ttcgtgccgg ccgagtccac
      961 gcgcgagttt cgcatctcgg tttcggactt tacgctctcc gtcctcatcc cccgggtgct
     1021 ggcgcgcgcg cacgccgagg gcaagcacat ccgctttgcc ctgatgccgc aggtgcaaga
     1081 cccgacccgc tcgctggatc gggccgaggt ggacctgctg gtcttgccgc aggaattctg
     1141 cacgcccgat catcctgccg aagaggtctt ccgcgaacgg catgtctgcg tggtctggcg
     1201 cgacagtgcg ctggcgcaag gcgagctgac gctggaacgc tacatggcct caggccatgt
     1261 ggtgatggtg ccgcctgggg ccaatgcgtc gtcggtggag gcgtggatgg ccaggaagct
     1321 gggctttgcg cgccgggtgg aagtgaccag cttcagcttc gcttctgcgc tggcgctggt
     1381 acaggggacg gaccgcatcg ccacggtgca tgcccggctg gcgcagctgc tggctccgca
     1441 atggccggtg gtgatcaagg agagtccgct gtcgctgggc gagatgcggc agatgatgca
     1501 gtggcatcgc taccgcagca atgatcctgg catccagtgg ctgcgtcggg tgtttctgga
     1561 gagtgcgcag gagatggatg cggcgctgcc aggcatctgc ggatccgatc caaaaaagaa
     1621 gagaaaggta gatccaaaaa agaagagaaa ggtataactc gagtggcatt gaattgaatt
     1681 gaaatcgata gatcaatttt tttcttttct ctttccccat cctttacgct aaaataatag
     1741 tttattttat tttttgaata ttttttattt atatacgtat atatagacta ttatttatct
     1801 tttaatgatt attaagattt ttattaaaaa aaaattcgct cctcttttaa tgcctttatg
     1861 cagttttttt ttcccattcg atatttctat gtgctgccaa accagatgtc aacacagcta
     1921 caacgaagct tcgtcttttc ttgaagaaaa gaggaagctc gctaaatggg attccacttt
     1981 ccgttccctg ccagctgatg gaaaaaggtt agtggaacga tgaagaataa aaagagagat
     2041 ccactgaggt gaaatttcag ctgacagcga gtttcatgat cgtgatgaac aatggtaacg
     2101 agttgtggct gttgccaggg agggtggttc tcaccaaatc gctacttggg tttgagattg
     2161 gtaaaattga ttgtttggat ggcaatcatc tatggaatgg atattatata acaaagaaga
     2221 aataatgaac tgattctctt cctccttctt gtcctttctt aattctgttg taattacctt
     2281 cctttgtaat tttttttgta attattcttc ttaataatcc aaacaaacac acatattaca
     2341 ataagatcta tggtttcaaa gggcgaagaa gacaatatgg ctattattaa ggaattcatg
     2401 agattcaagg tccacatgga aggttctgtc aacggtcacg aattcgaaat tgaaggtgaa
     2461 ggtgaaggta gaccatacga aggtacccaa accgctaagt tgaaggtcac caagggtggt
     2521 ccattgccat tcgcttggga tattttgtct ccacaattca tgtacggttc taaggcttac
     2581 gtcaagcacc cagctgatat tccagattac ttgaagttgt ctttcccaga aggtttcaag
     2641 tgggaaagag tcatgaactt cgaagatggt ggtgtcgtca ccgtcaccca agattcttct
     2701 ttgcaagatg gtgaattcat ttacaaggtc aagttgagag gtaccaactt cccatctgat
     2761 ggtccagtca tgcaaaagaa gaccatgggt tgggaagctt cttctgaaag aatgtaccca
     2821 gaagatggtg ctttgaaggg tgaaattaag caaagattga agttgaagga tggtggtcac
     2881 tacgatgctg aagtcaagac cacctacaag gctaagaagc cagtccaatt gccaggtgct
     2941 tacaacgtca acattaagtt ggatattacc tctcacaacg aagattacac cattgtcgaa
     3001 caatacgaaa gagctgaagg tagacactct accggtggta tggatgaatt gtacaagtaa
     3061 atcctaactc gagataaagc aatcttgatg aggataatga tttttttttg aatatacata
     3121 aatactaccg tttttctgct agattttgtg atgacgtaaa taagtacata ttacttttta
     3181 agccaagaca agattaagca ttaactttac ccttttcttt ctaagtttca atattagtta
     3241 tcactgttta aaagttatgg cgagaacgtc ggcggttaaa atatattacc ctgaacggct
     3301 gagcagttac agagatgtta cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     3361 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     3421 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     3481 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     3541 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     3601 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     3661 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     3721 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     3781 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     3841 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     3901 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     3961 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     4021 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     4081 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     4141 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     4201 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     4261 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     4321 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     4381 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     4441 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     4501 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     4561 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     4621 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     4681 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     4741 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     4801 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     4861 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     4921 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     4981 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     5041 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     5101 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     5161 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     5221 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     5281 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     5341 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     5401 aggaacttcc cgagcggccg cgtgttacaa ccaattaacc aattctgatt agaaaaactc
     5461 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     5521 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     5581 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     5641 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     5701 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     5761 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     5821 gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     5881 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     5941 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     6001 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     6061 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     6121 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     6181 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga
     6241 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     6301 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     6361 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agtcatgacc
     6421 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6481 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6541 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6601 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6661 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6721 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6781 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6841 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6901 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6961 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7021 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7081 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7141 tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg ccgcccctga attcgcatct
     7201 agactgaact ggccgataat tgcagacgaa cgcagttcga gttta
//
