LOCUS       sequence-258372-        6748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pRPL18B_NLS_FdeR-N_tPGK1-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6748)
  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 6748)
  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_002_026_MV1
FEATURES             Location/Qualifiers
     source          1..6748
                     /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             2533..2553
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5355..6158
                     /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    6241..6744
                     /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 tgccaaagaa gaagagaaag gttggttctc gtttcaacaa gctcgacctc
     2581 aatcttctgg tcgccctgga tgcactgctc acggagatga gcatcagccg cgccgccgaa
     2641 aagatccatc tgagccagtc ggccatgagc aatgccctgg cgcggctgcg cgagtatttc
     2701 gatgatgaat tgctgatcca ggtgggccgg cgcatggagc ccacgccgcg cgccgaggtg
     2761 ctcaaggatg cggtgcatga tgtgctgcgg cgtatcgatg gctccatcgc ggcgctgccg
     2821 gccttcgtgc cggccgagtc cacgcgcgag tttcgcatct cggtttcgga ctttacgctc
     2881 tccgtcctca tcccccgggt gctggcgcgc gcgcacgccg agggcaagca catccgcttt
     2941 gccctgatgc cgcaggtgca agacccgacc cgctcgctgg atcgggccga ggtggacctg
     3001 ctggtcttgc cgcaggaatt ctgcacgccc gatcatcctg ccgaagaggt cttccgcgaa
     3061 cggcatgtct gcgtggtctg gcgcgacagt gcgctggcgc aaggcgagct gacgctggaa
     3121 cgctacatgg cctcaggcca tgtggtgatg gtgccgcctg gggccaatgc gtcgtcggtg
     3181 gaggcgtgga tggccaggaa gctgggcttt gcgcgccggg tggaagtgac cagcttcagc
     3241 ttcgcttctg cgctggcgct ggtacagggg acggaccgca tcgccacggt gcatgcccgg
     3301 ctggcgcagc tgctggctcc gcaatggccg gtggtgatca aggagagtcc gctgtcgctg
     3361 ggcgagatgc ggcagatgat gcagtggcat cgctaccgca gcaatgatcc tggcatccag
     3421 tggctgcgtc gggtgtttct ggagagtgcg caggagatgg atgcggcgct gccaggcatc
     3481 tgctgaatcc taactcgaga ttgaattgaa ttgaaatcga tagatcaatt tttttctttt
     3541 ctctttcccc atcctttacg ctaaaataat agtttatttt attttttgaa tattttttat
     3601 ttatatacgt atatatagac tattatttat cttttaatga ttattaagat ttttattaaa
     3661 aaaaaattcg ctcctctttt aatgccttta tgcagttttt ttttcccatt cgatatttct
     3721 atgtgctgcc aaaccagatg tcaacacagc tacaacgaag cttcgtcttt tcttgaagaa
     3781 aagaggaagc tcgctaaatg ggattccact ttccgttccc tgccagctga tggaaaaagg
     3841 ttagtggaac gatgaagaat aaaaagagag atccactgag gtgaaatttc agctgacagc
     3901 gagtttcatg atcgtgatga acaatggtaa cgagttgtgg ctgttgccag ggagggtggt
     3961 tctcaccaaa tcgctacttg ggtttgagat tggtaaaatt gattgtttgg atggcaatca
     4021 tctatggaat ggatattata taacaaagaa gaaataatga actgattctc ttcctccttc
     4081 ttgtcctttc ttaattctgt tgtaattacc ttcctttgta attttttttg taattattct
     4141 tcttaataat ccaaacaaac acacatatta caataagatc tatggtttca aagggcgaag
     4201 aagacaatat ggctattatt aaggaattca tgagattcaa ggtccacatg gaaggttctg
     4261 tcaacggtca cgaattcgaa attgaaggtg aaggtgaagg tagaccatac gaaggtaccc
     4321 aaaccgctaa gttgaaggtc accaagggtg gtccattgcc attcgcttgg gatattttgt
     4381 ctccacaatt catgtacggt tctaaggctt acgtcaagca cccagctgat attccagatt
     4441 acttgaagtt gtctttccca gaaggtttca agtgggaaag agtcatgaac ttcgaagatg
     4501 gtggtgtcgt caccgtcacc caagattctt ctttgcaaga tggtgaattc atttacaagg
     4561 tcaagttgag aggtaccaac ttcccatctg atggtccagt catgcaaaag aagaccatgg
     4621 gttgggaagc ttcttctgaa agaatgtacc cagaagatgg tgctttgaag ggtgaaatta
     4681 agcaaagatt gaagttgaag gatggtggtc actacgatgc tgaagtcaag accacctaca
     4741 aggctaagaa gccagtccaa ttgccaggtg cttacaacgt caacattaag ttggatatta
     4801 cctctcacaa cgaagattac accattgtcg aacaatacga aagagctgaa ggtagacact
     4861 ctaccggtgg tatggatgaa ttgtacaagt aaatcctaac tcgagataaa gcaatcttga
     4921 tgaggataat gatttttttt tgaatataca taaatactac cgtttttctg ctagattttg
     4981 tgatgacgta aataagtaca tattactttt taagccaaga caagattaag cattaacttt
     5041 acccttttct ttctaagttt caatattagt tatcactgtt taaaagttat ggcgagaacg
     5101 tcggcggtta aaatatatta ccctgaacgg ctgagcagtt acagagatgt tacgaaccac
     5161 tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     5221 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     5281 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acgtgcagga
     5341 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     5401 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     5461 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     5521 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     5581 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg
     5641 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     5701 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     5761 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     5821 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     5881 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     5941 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac
     6001 agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag
     6061 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     6121 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     6181 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
     6241 atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa
     6301 atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag
     6361 actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat
     6421 taatgccgaa ttgtttcatc ttgtctgtgt agaaaaccac acacgaaaat cctgtgattt
     6481 tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata
     6541 aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt
     6601 cttcattccg taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat
     6661 aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg
     6721 cgtgtaagtt acaggcaagc gatccgtc
//
