LOCUS       sequence-258380-        7221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV2-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 7221)
  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 7221)
  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_MV2
FEATURES             Location/Qualifiers
     source          1..7221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(40..855)
                     /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(40..855)
                     /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     763..782
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     847..866
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1080..1668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1569..1588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1837..2503
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2373..2396
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2513..2533
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5335..6138
                     /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      6221..7203
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ccgagcggcc gcgtgttaca accaattaac caattctgat tagaaaaact catcgagcat
       61 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
      121 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
      181 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
      241 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
      301 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
      361 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac
      421 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
      481 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
      541 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
      601 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
      661 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
      721 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
      781 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg
      841 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
      901 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
      961 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagtcatgac caaaatccct
     1021 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1741 ttatcccctg attctgtgga taaccgtgcg gccgcccctg aattcgcatc tagactgaac
     1801 tggccgataa ttgcagacga acgcagttcg agtttatcat tatcaatact gccatttcaa
     1861 agaatacgta aataattaat agtagtgatt ttcctaactt tatttagtca aaaaattagc
     1921 cttttaattc tgctgtaacc cgtacatgcc caaaataggg ggcgggttac acagaatata
     1981 taacatcgta ggtgtctggg tgaacagttt attcctggca tccactaaat ataatggagc
     2041 ccgcttttta agctggcatc cagaaaaaaa aagaatccca gcaccaaaat attgttttct
     2101 tcaccaacca tcagttcata ggtccattct cttagcgcaa ctacagagaa caggggcaca
     2161 aacaggcaaa aaacgggcac aacctcaatg gagtgatgca acctgcctgg agtaaatgat
     2221 gacacaaggc aattgaccca cgcatgtatc tatctcattt tcttacacct tctattacct
     2281 tctgctctct ctgatttgga aaaagctgaa aaaaaaggtt gaaaccagtt ccctgaaatt
     2341 attcccctac ttgactaata agtatataaa gacggtaggt attgattgta attctgtaaa
     2401 tctatttctt aaacttctta aattctactt ttatagttag tctttttttt agttttaaaa
     2461 caccaagaac ttagtttcga ataaacacac ataaacaaac aaaagatcta tgccaaagaa
     2521 gaagagaaag gttggttctc gtttcaacaa gctcgacctc aatcttctgg tcgccctgga
     2581 tgcactgctc acggagatga gcatcagccg cgccgccgaa aagatccatc tgagccagtc
     2641 ggccatgagc aatgccctgg cgcggctgcg cgagtatttc gatgatgaat tgctgatcca
     2701 ggtgggccgg cgcatggagc ccacgccgcg cgccgaggtg ctcaaggatg cggtgcatga
     2761 tgtgctgcgg cgtatcgatg gctccatcgc ggcgctgccg gccttcgtgc cggccgagtc
     2821 cacgcgcgag tttcgcatct cggtttcgga ctttacgctc tccgtcctca tcccccgggt
     2881 gctggcgcgc gcgcacgccg agggcaagca catccgcttt gccctgatgc cgcaggtgca
     2941 agacccgacc cgctcgctgg atcgggccga ggtggacctg ctggtcttgc cgcaggaatt
     3001 ctgcacgccc gatcatcctg ccgaagaggt cttccgcgaa cggcatgtct gcgtggtctg
     3061 gcgcgacagt gcgctggcgc aaggcgagct gacgctggaa cgctacatgg cctcaggcca
     3121 tgtggtgatg gtgccgcctg gggccaatgc gtcgtcggtg gaggcgtgga tggccaggaa
     3181 gctgggcttt gcgcgccggg tggaagtgac cagcttcagc ttcgcttctg cgctggcgct
     3241 ggtacagggg acggaccgca tcgccacggt gcatgcccgg ctggcgcagc tgctggctcc
     3301 gcaatggccg gtggtgatca aggagagtcc gctgtcgctg ggcgagatgc ggcagatgat
     3361 gcagtggcat cgctaccgca gcaatgatcc tggcatccag tggctgcgtc gggtgtttct
     3421 ggagagtgcg caggagatgg atgcggcgct gccaggcatc tgctgaatcc taactcgaga
     3481 ttgaattgaa ttgaaatcga tagatcaatt tttttctttt ctctttcccc atcctttacg
     3541 ctaaaataat agtttatttt attttttgaa tattttttat ttatatacgt atatatagac
     3601 tattatttat cttttaatga ttattaagat ttttattaaa aaaaaattcg ctcctctttt
     3661 aatgccttta tgcagttttt ttttcccatt cgatatttct atgtgctgcc aaaccagatg
     3721 tcaacacagc tacaacgaag cttcgtcttt tcttgaagaa aagaggaagc tcgctaaatg
     3781 ggattccact ttccgttccc tgccagctga tggaaaaagg ttagtggaac gatgaagaat
     3841 aaaaagagag atccactgag gtgaaatttc agctgacagc gagtttcatg atcgtgatga
     3901 acaatggtaa cgagttgtgg ctgttgccag ggagggtggt tctcaccaaa tcgctacttg
     3961 ggtttgagat tggtaaaatt gattgtttgg atggcaatca tctatggaat ggatattata
     4021 taacaaagaa gaaataatga actgattctc ttcctccttc ttgtcctttc ttaattctgt
     4081 tgtaattacc ttcctttgta attttttttg taattattct tcttaataat ccaaacaaac
     4141 acacatatta caataagatc tatggtttca aagggcgaag aagacaatat ggctattatt
     4201 aaggaattca tgagattcaa ggtccacatg gaaggttctg tcaacggtca cgaattcgaa
     4261 attgaaggtg aaggtgaagg tagaccatac gaaggtaccc aaaccgctaa gttgaaggtc
     4321 accaagggtg gtccattgcc attcgcttgg gatattttgt ctccacaatt catgtacggt
     4381 tctaaggctt acgtcaagca cccagctgat attccagatt acttgaagtt gtctttccca
     4441 gaaggtttca agtgggaaag agtcatgaac ttcgaagatg gtggtgtcgt caccgtcacc
     4501 caagattctt ctttgcaaga tggtgaattc atttacaagg tcaagttgag aggtaccaac
     4561 ttcccatctg atggtccagt catgcaaaag aagaccatgg gttgggaagc ttcttctgaa
     4621 agaatgtacc cagaagatgg tgctttgaag ggtgaaatta agcaaagatt gaagttgaag
     4681 gatggtggtc actacgatgc tgaagtcaag accacctaca aggctaagaa gccagtccaa
     4741 ttgccaggtg cttacaacgt caacattaag ttggatatta cctctcacaa cgaagattac
     4801 accattgtcg aacaatacga aagagctgaa ggtagacact ctaccggtgg tatggatgaa
     4861 ttgtacaagt aaatcctaac tcgagataaa gcaatcttga tgaggataat gatttttttt
     4921 tgaatataca taaatactac cgtttttctg ctagattttg tgatgacgta aataagtaca
     4981 tattactttt taagccaaga caagattaag cattaacttt acccttttct ttctaagttt
     5041 caatattagt tatcactgtt taaaagttat ggcgagaacg tcggcggtta aaatatatta
     5101 ccctgaacgg ctgagcagtt acagagatgt tacgaaccac tagtgcactg cagtacacgg
     5161 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     5221 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     5281 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     5341 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     5401 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     5461 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     5521 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     5581 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     5641 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     5701 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     5761 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     5821 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     5881 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     5941 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     6001 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     6061 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     6121 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     6181 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     6241 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     6301 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     6361 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     6421 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     6481 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     6541 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     6601 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     6661 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     6721 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     6781 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     6841 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     6901 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     6961 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     7021 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     7081 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     7141 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     7201 tagctagaga ataggaactt c
//
