LOCUS       sequence-266898-        6830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pRPL18B_ NLS_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 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_008_026_MV1
FEATURES             Location/Qualifiers
     source          1..6830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..21
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             992..1012
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1016..1036
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2905..3708
                     /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    3791..4294
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4338..5153)
                     /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(4338..5153)
                     /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     5061..5080
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5145..5164
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5378..5966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5867..5886
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaaagaaga agagaaaggt tggttctcgt ttcaacaagc tcgacctcaa tcttctggtc
       61 gccctggatg cactgctcac ggagatgagc atcagccgcg ccgccgaaaa gatccatctg
      121 agccagtcgg ccatgagcaa tgccctggcg cggctgcgcg agtatttcga tgatgaattg
      181 ctgatccagg tgggccggcg catggagccc acgccgcgcg ccgaggtgct caaggatgcg
      241 gtgcatgatg tgctgcggcg tatcgatggc tccatcgcgg cgctgccggc cttcgtgccg
      301 gccgagtcca cgcgcgagtt tcgcatctcg gtttcggact ttacgctctc cgtcctcatc
      361 ccccgggtgc tggcgcgcgc gcacgccgag ggcaagcaca tccgctttgc cctgatgccg
      421 caggtgcaag acccgacccg ctcgctggat cgggccgagg tggacctgct ggtcttgccg
      481 caggaattct gcacgcccga tcatcctgcc gaagaggtct tccgcgaacg gcatgtctgc
      541 gtggtctggc gcgacagtgc gctggcgcaa ggcgagctga cgctggaacg ctacatggcc
      601 tcaggccatg tggtgatggt gccgcctggg gccaatgcgt cgtcggtgga ggcgtggatg
      661 gccaggaagc tgggctttgc gcgccgggtg gaagtgacca gcttcagctt cgcttctgcg
      721 ctggcgctgg tacaggggac ggaccgcatc gccacggtgc atgcccggct ggcgcagctg
      781 ctggctccgc aatggccggt ggtgatcaag gagagtccgc tgtcgctggg cgagatgcgg
      841 cagatgatgc agtggcatcg ctaccgcagc aatgatcctg gcatccagtg gctgcgtcgg
      901 gtgtttctgg agagtgcgca ggagatggat gcggcgctgc caggcatctg cggatcctga
      961 gaccaggctg ccaggcatct gcggatccga tccaaaaaag aagagaaagg tagatccaaa
     1021 aaagaagaga aaggtataac tcgagtggca ttgaattgaa ttgaaatcga tagatcaatt
     1081 tttttctttt ctctttcccc atcctttacg ctaaaataat agtttatttt attttttgaa
     1141 tattttttat ttatatacgt atatatagac tattatttat cttttaatga ttattaagat
     1201 ttttattaaa aaaaaattcg ctcctctttt aatgccttta tgcagttttt ttttcccatt
     1261 cgatatttct atgtgctgcc aaaccagatg tcaacacagc tacaacgaag cttcgtcttt
     1321 tcttgaagaa aagaggaagc tcgctaaatg ggattccact ttccgttccc tgccagctga
     1381 tggaaaaagg ttagtggaac gatgaagaat aaaaagagag atccactgag gtgaaatttc
     1441 agctgacagc gagtttcatg atcgtgatga acaatggtaa cgagttgtgg ctgttgccag
     1501 ggagggtggt tctcaccaaa tcgctacttg ggtttgagat tggtaaaatt gattgtttgg
     1561 atggcaatca tctatggaat ggatattata taacaaagaa gaaataatga actgattctc
     1621 ttcctccttc ttgtcctttc ttaattctgt tgtaattacc ttcctttgta attttttttg
     1681 taattattct tcttaataat ccaaacaaac acacatatta caataagatc tatggtttca
     1741 aagggcgaag aagacaatat ggctattatt aaggaattca tgagattcaa ggtccacatg
     1801 gaaggttctg tcaacggtca cgaattcgaa attgaaggtg aaggtgaagg tagaccatac
     1861 gaaggtaccc aaaccgctaa gttgaaggtc accaagggtg gtccattgcc attcgcttgg
     1921 gatattttgt ctccacaatt catgtacggt tctaaggctt acgtcaagca cccagctgat
     1981 attccagatt acttgaagtt gtctttccca gaaggtttca agtgggaaag agtcatgaac
     2041 ttcgaagatg gtggtgtcgt caccgtcacc caagattctt ctttgcaaga tggtgaattc
     2101 atttacaagg tcaagttgag aggtaccaac ttcccatctg atggtccagt catgcaaaag
     2161 aagaccatgg gttgggaagc ttcttctgaa agaatgtacc cagaagatgg tgctttgaag
     2221 ggtgaaatta agcaaagatt gaagttgaag gatggtggtc actacgatgc tgaagtcaag
     2281 accacctaca aggctaagaa gccagtccaa ttgccaggtg cttacaacgt caacattaag
     2341 ttggatatta cctctcacaa cgaagattac accattgtcg aacaatacga aagagctgaa
     2401 ggtagacact ctaccggtgg tatggatgaa ttgtacaagt aaatcctaac tcgagataaa
     2461 gcaatcttga tgaggataat gatttttttt tgaatataca taaatactac cgtttttctg
     2521 ctagattttg tgatgacgta aataagtaca tattactttt taagccaaga caagattaag
     2581 cattaacttt acccttttct ttctaagttt caatattagt tatcactgtt taaaagttat
     2641 ggcgagaacg tcggcggtta aaatatatta ccctgaacgg ctgagcagtt acagagatgt
     2701 tacgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     2761 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     2821 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     2881 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     2941 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     3001 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     3061 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     3121 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     3181 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     3241 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     3301 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     3361 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     3421 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     3481 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     3541 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     3601 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     3661 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     3721 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     3781 ttacccgagt atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata
     3841 taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa
     3901 gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc
     3961 tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaaaaccac acacgaaaat
     4021 cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc
     4081 ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt
     4141 attttttttt cttcattccg taactcttct accttcttta tttactttct aaaatccaaa
     4201 tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag
     4261 atacgaggcg cgtgtaagtt acaggcaagc gatccgtccc gagcggccgc gtgttacaac
     4321 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     4381 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     4441 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     4501 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     4561 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag
     4621 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     4681 ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa
     4741 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     4801 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     4861 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     4921 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     4981 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     5041 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     5101 ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct cataacaccc
     5161 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     5221 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     5281 tttgctgagt tgaaggatca gtcatgacca aaatccctta acgtgagttt tcgttccact
     5341 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5401 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5461 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5521 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5581 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5641 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5701 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5761 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5821 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5881 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5941 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6001 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6061 accgtgcggc cgcccctgaa ttcgcatcta gactgaactg gccgataatt gcagacgaac
     6121 gaagaggatg tccaatattt tttttaagga ataaggatac ttcaagacta gattcccccc
     6181 tgcattccca tcagaaccgt aaaccttggc gctttccttg ggaagtattc aagaagtgcc
     6241 ttgtccggtt tctgtggctc acaaaccagc gcgcccgata tggctttctt ttcacttatg
     6301 aatgtaccag tacgggacaa ttagaacgct cctgtaacaa tctctttgca aatgtggggt
     6361 tacattctaa ccatgtcaca ctgctgacga aattcaaagt aaaaaaaaat gggaccacgt
     6421 cttgagaacg atagattttc tttattttac attgaacagt cgttgtctca gcgcgcttta
     6481 tgttttcatt catacttcat attataaaat aacaaaagaa gaatttcata ttcacgccca
     6541 agaaatcagg ctgctttcca aatgcaattg acacttcatt agccatcaca caaaactctt
     6601 tcttgctgga gcttctttta aaaaagacct cagtacacca aacacgttac ccgacctcgt
     6661 tattttacga caactatgat aaaattctga agaaaaaata aaaaaatttt catacttctt
     6721 gcttttattt aaaccattga atgatttctt ttgaacaaaa ctacctgttt caccaaagga
     6781 aatagaaaga aaaaatcaat tagaagaaaa caaaaaacaa aagatctatg
//
