LOCUS       sequence-266876-        6830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pREV1_ NLS_FdeR-N_2NLS-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6830)
  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 6830)
  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_009_026_MV1
FEATURES             Location/Qualifiers
     source          1..6830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(506..1321)
                     /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(506..1321)
                     /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     1229..1248
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1313..1332
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1546..2134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2035..2054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2999..3019
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3990..4010
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4014..4034
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5903..6706
                     /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"
     misc_feature    join(6789..6830,1..462)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
       61 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
      121 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aaaaccacac
      181 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
      241 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
      301 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
      361 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
      421 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcccga gcggccgcgt
      481 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
      541 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
      601 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
      661 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
      721 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
      781 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
      841 ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa
      901 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
      961 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     1021 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     1081 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     1141 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     1201 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     1261 catccatgtt ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca
     1321 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     1381 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     1441 atcgaacttt tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagttttc
     1501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1681 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2221 tgtggataac cgtgcggccg cccctgaatt cgcatctaga ctgaactggc cgataattgc
     2281 agacgaacgg tgttgttatc cgatacaacc ggatattttt cttttaatga gtctaaaccg
     2341 tgatagcttc aggttaatac aatcaaaaaa agctcaaata ttcttttaat gccgcgttca
     2401 cagattccaa ttgaatacaa ctaggtagtt cattatatga agcctttgct actatttttc
     2461 actatagtct gccttcacct taatgcagac atccacatat tttaatcact ttaaaataaa
     2521 aaggaagata tattagaagc tatgatccaa tctgtaagcc agattaaaat tcacgaactc
     2581 ttctttcatt tgaattgaat gctttgagtt ggggtagatt atcgcaaatt actcatcaca
     2641 tttattgact acgaacttgc tgatgtcctt tttttattta tatttttctt cagtgaagcg
     2701 attttttttt tacacagacc aagacggaaa aaagtagcta aggaagaaaa caaaatcatg
     2761 aaaaaaatgt gaagtgatca tgcacatcgc atcaacttaa acattggctt agagatatat
     2821 agagttagag tttacggcaa cctttaagca ccaatacctt ttggcatagt ctaaagacct
     2881 ggttcttaat tttaaacaaa tttaactaaa gatttcccta tcaaagaagt aacgagttga
     2941 cagattttct caaaataaat cgatactgca tttctaggca tatccagcga gatctatgcc
     3001 aaagaagaag agaaaggttg gttctcgttt caacaagctc gacctcaatc ttctggtcgc
     3061 cctggatgca ctgctcacgg agatgagcat cagccgcgcc gccgaaaaga tccatctgag
     3121 ccagtcggcc atgagcaatg ccctggcgcg gctgcgcgag tatttcgatg atgaattgct
     3181 gatccaggtg ggccggcgca tggagcccac gccgcgcgcc gaggtgctca aggatgcggt
     3241 gcatgatgtg ctgcggcgta tcgatggctc catcgcggcg ctgccggcct tcgtgccggc
     3301 cgagtccacg cgcgagtttc gcatctcggt ttcggacttt acgctctccg tcctcatccc
     3361 ccgggtgctg gcgcgcgcgc acgccgaggg caagcacatc cgctttgccc tgatgccgca
     3421 ggtgcaagac ccgacccgct cgctggatcg ggccgaggtg gacctgctgg tcttgccgca
     3481 ggaattctgc acgcccgatc atcctgccga agaggtcttc cgcgaacggc atgtctgcgt
     3541 ggtctggcgc gacagtgcgc tggcgcaagg cgagctgacg ctggaacgct acatggcctc
     3601 aggccatgtg gtgatggtgc cgcctggggc caatgcgtcg tcggtggagg cgtggatggc
     3661 caggaagctg ggctttgcgc gccgggtgga agtgaccagc ttcagcttcg cttctgcgct
     3721 ggcgctggta caggggacgg accgcatcgc cacggtgcat gcccggctgg cgcagctgct
     3781 ggctccgcaa tggccggtgg tgatcaagga gagtccgctg tcgctgggcg agatgcggca
     3841 gatgatgcag tggcatcgct accgcagcaa tgatcctggc atccagtggc tgcgtcgggt
     3901 gtttctggag agtgcgcagg agatggatgc ggcgctgcca ggcatctgcg gatcctgaga
     3961 ccaggctgcc aggcatctgc ggatccgatc caaaaaagaa gagaaaggta gatccaaaaa
     4021 agaagagaaa ggtataactc gagtggcatt gaattgaatt gaaatcgata gatcaatttt
     4081 tttcttttct ctttccccat cctttacgct aaaataatag tttattttat tttttgaata
     4141 ttttttattt atatacgtat atatagacta ttatttatct tttaatgatt attaagattt
     4201 ttattaaaaa aaaattcgct cctcttttaa tgcctttatg cagttttttt ttcccattcg
     4261 atatttctat gtgctgccaa accagatgtc aacacagcta caacgaagct tcgtcttttc
     4321 ttgaagaaaa gaggaagctc gctaaatggg attccacttt ccgttccctg ccagctgatg
     4381 gaaaaaggtt agtggaacga tgaagaataa aaagagagat ccactgaggt gaaatttcag
     4441 ctgacagcga gtttcatgat cgtgatgaac aatggtaacg agttgtggct gttgccaggg
     4501 agggtggttc tcaccaaatc gctacttggg tttgagattg gtaaaattga ttgtttggat
     4561 ggcaatcatc tatggaatgg atattatata acaaagaaga aataatgaac tgattctctt
     4621 cctccttctt gtcctttctt aattctgttg taattacctt cctttgtaat tttttttgta
     4681 attattcttc ttaataatcc aaacaaacac acatattaca ataagatcta tggtttcaaa
     4741 gggcgaagaa gacaatatgg ctattattaa ggaattcatg agattcaagg tccacatgga
     4801 aggttctgtc aacggtcacg aattcgaaat tgaaggtgaa ggtgaaggta gaccatacga
     4861 aggtacccaa accgctaagt tgaaggtcac caagggtggt ccattgccat tcgcttggga
     4921 tattttgtct ccacaattca tgtacggttc taaggcttac gtcaagcacc cagctgatat
     4981 tccagattac ttgaagttgt ctttcccaga aggtttcaag tgggaaagag tcatgaactt
     5041 cgaagatggt ggtgtcgtca ccgtcaccca agattcttct ttgcaagatg gtgaattcat
     5101 ttacaaggtc aagttgagag gtaccaactt cccatctgat ggtccagtca tgcaaaagaa
     5161 gaccatgggt tgggaagctt cttctgaaag aatgtaccca gaagatggtg ctttgaaggg
     5221 tgaaattaag caaagattga agttgaagga tggtggtcac tacgatgctg aagtcaagac
     5281 cacctacaag gctaagaagc cagtccaatt gccaggtgct tacaacgtca acattaagtt
     5341 ggatattacc tctcacaacg aagattacac cattgtcgaa caatacgaaa gagctgaagg
     5401 tagacactct accggtggta tggatgaatt gtacaagtaa atcctaactc gagataaagc
     5461 aatcttgatg aggataatga tttttttttg aatatacata aatactaccg tttttctgct
     5521 agattttgtg atgacgtaaa taagtacata ttacttttta agccaagaca agattaagca
     5581 ttaactttac ccttttcttt ctaagtttca atattagtta tcactgttta aaagttatgg
     5641 cgagaacgtc ggcggttaaa atatattacc ctgaacggct gagcagttac agagatgtta
     5701 cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc
     5761 cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg
     5821 tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac
     5881 gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca
     5941 tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg
     6001 tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc
     6061 caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac
     6121 agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa
     6181 atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc
     6241 agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt
     6301 gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt
     6361 gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa
     6421 gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga
     6481 aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt
     6541 gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt
     6601 tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
     6661 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
     6721 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
     6781 acccgagtat cacgtgctat aaaaataatt ataatttaaa ttttttaata
//
