LOCUS       sequence-258375-        6772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pRPL18B_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 6772)
  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 6772)
  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_005_026_MV1
FEATURES             Location/Qualifiers
     source          1..6772
                     /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"
     CDS             3466..3486
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3490..3510
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5379..6182
                     /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    6265..6768
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
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 acgaagagga tgtccaatat tttttttaag gaataaggat
     1861 acttcaagac tagattcccc cctgcattcc catcagaacc gtaaaccttg gcgctttcct
     1921 tgggaagtat tcaagaagtg ccttgtccgg tttctgtggc tcacaaacca gcgcgcccga
     1981 tatggctttc ttttcactta tgaatgtacc agtacgggac aattagaacg ctcctgtaac
     2041 aatctctttg caaatgtggg gttacattct aaccatgtca cactgctgac gaaattcaaa
     2101 gtaaaaaaaa atgggaccac gtcttgagaa cgatagattt tctttatttt acattgaaca
     2161 gtcgttgtct cagcgcgctt tatgttttca ttcatacttc atattataaa ataacaaaag
     2221 aagaatttca tattcacgcc caagaaatca ggctgctttc caaatgcaat tgacacttca
     2281 ttagccatca cacaaaactc tttcttgctg gagcttcttt taaaaaagac ctcagtacac
     2341 caaacacgtt acccgacctc gttattttac gacaactatg ataaaattct gaagaaaaaa
     2401 taaaaaaatt ttcatacttc ttgcttttat ttaaaccatt gaatgatttc ttttgaacaa
     2461 aactacctgt ttcaccaaag gaaatagaaa gaaaaaatca attagaagaa aacaaaaaac
     2521 aaaagatcta tgcgtttcaa caagctcgac ctcaatcttc tggtcgccct ggatgcactg
     2581 ctcacggaga tgagcatcag ccgcgccgcc gaaaagatcc atctgagcca gtcggccatg
     2641 agcaatgccc tggcgcggct gcgcgagtat ttcgatgatg aattgctgat ccaggtgggc
     2701 cggcgcatgg agcccacgcc gcgcgccgag gtgctcaagg atgcggtgca tgatgtgctg
     2761 cggcgtatcg atggctccat cgcggcgctg ccggccttcg tgccggccga gtccacgcgc
     2821 gagtttcgca tctcggtttc ggactttacg ctctccgtcc tcatcccccg ggtgctggcg
     2881 cgcgcgcacg ccgagggcaa gcacatccgc tttgccctga tgccgcaggt gcaagacccg
     2941 acccgctcgc tggatcgggc cgaggtggac ctgctggtct tgccgcagga attctgcacg
     3001 cccgatcatc ctgccgaaga ggtcttccgc gaacggcatg tctgcgtggt ctggcgcgac
     3061 agtgcgctgg cgcaaggcga gctgacgctg gaacgctaca tggcctcagg ccatgtggtg
     3121 atggtgccgc ctggggccaa tgcgtcgtcg gtggaggcgt ggatggccag gaagctgggc
     3181 tttgcgcgcc gggtggaagt gaccagcttc agcttcgctt ctgcgctggc gctggtacag
     3241 gggacggacc gcatcgccac ggtgcatgcc cggctggcgc agctgctggc tccgcaatgg
     3301 ccggtggtga tcaaggagag tccgctgtcg ctgggcgaga tgcggcagat gatgcagtgg
     3361 catcgctacc gcagcaatga tcctggcatc cagtggctgc gtcgggtgtt tctggagagt
     3421 gcgcaggaga tggatgcggc gctgccaggc atctgcggat ccgatccaaa aaagaagaga
     3481 aaggtagatc caaaaaagaa gagaaaggta taactcgagt ggcattgaat tgaattgaaa
     3541 tcgatagatc aatttttttc ttttctcttt ccccatcctt tacgctaaaa taatagttta
     3601 ttttattttt tgaatatttt ttatttatat acgtatatat agactattat ttatctttta
     3661 atgattatta agatttttat taaaaaaaaa ttcgctcctc ttttaatgcc tttatgcagt
     3721 ttttttttcc cattcgatat ttctatgtgc tgccaaacca gatgtcaaca cagctacaac
     3781 gaagcttcgt cttttcttga agaaaagagg aagctcgcta aatgggattc cactttccgt
     3841 tccctgccag ctgatggaaa aaggttagtg gaacgatgaa gaataaaaag agagatccac
     3901 tgaggtgaaa tttcagctga cagcgagttt catgatcgtg atgaacaatg gtaacgagtt
     3961 gtggctgttg ccagggaggg tggttctcac caaatcgcta cttgggtttg agattggtaa
     4021 aattgattgt ttggatggca atcatctatg gaatggatat tatataacaa agaagaaata
     4081 atgaactgat tctcttcctc cttcttgtcc tttcttaatt ctgttgtaat taccttcctt
     4141 tgtaattttt tttgtaatta ttcttcttaa taatccaaac aaacacacat attacaataa
     4201 gatctatggt ttcaaagggc gaagaagaca atatggctat tattaaggaa ttcatgagat
     4261 tcaaggtcca catggaaggt tctgtcaacg gtcacgaatt cgaaattgaa ggtgaaggtg
     4321 aaggtagacc atacgaaggt acccaaaccg ctaagttgaa ggtcaccaag ggtggtccat
     4381 tgccattcgc ttgggatatt ttgtctccac aattcatgta cggttctaag gcttacgtca
     4441 agcacccagc tgatattcca gattacttga agttgtcttt cccagaaggt ttcaagtggg
     4501 aaagagtcat gaacttcgaa gatggtggtg tcgtcaccgt cacccaagat tcttctttgc
     4561 aagatggtga attcatttac aaggtcaagt tgagaggtac caacttccca tctgatggtc
     4621 cagtcatgca aaagaagacc atgggttggg aagcttcttc tgaaagaatg tacccagaag
     4681 atggtgcttt gaagggtgaa attaagcaaa gattgaagtt gaaggatggt ggtcactacg
     4741 atgctgaagt caagaccacc tacaaggcta agaagccagt ccaattgcca ggtgcttaca
     4801 acgtcaacat taagttggat attacctctc acaacgaaga ttacaccatt gtcgaacaat
     4861 acgaaagagc tgaaggtaga cactctaccg gtggtatgga tgaattgtac aagtaaatcc
     4921 taactcgaga taaagcaatc ttgatgagga taatgatttt tttttgaata tacataaata
     4981 ctaccgtttt tctgctagat tttgtgatga cgtaaataag tacatattac tttttaagcc
     5041 aagacaagat taagcattaa ctttaccctt ttctttctaa gtttcaatat tagttatcac
     5101 tgtttaaaag ttatggcgag aacgtcggcg gttaaaatat attaccctga acggctgagc
     5161 agttacagag atgttacgaa ccactagtgc actgcagtac acggtttcct tgaaattttt
     5221 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     5281 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     5341 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     5401 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     5461 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     5521 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     5581 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     5641 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     5701 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     5761 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     5821 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     5881 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     5941 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     6001 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat
     6061 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     6121 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     6181 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     6241 aattatatca gttattaccc gagtatcacg tgctataaaa ataattataa tttaaatttt
     6301 ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt
     6361 ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat
     6421 cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaaa
     6481 ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat
     6541 ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt
     6601 ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact
     6661 ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat
     6721 tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg tc
//
