LOCUS       sequence-258378-        6799 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 6799)
  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 6799)
  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..6799
                     /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             3493..3513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3517..3537
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5406..6209
                     /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    6292..6795
                     /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 tgcggatccg atccaaaaaa gaagagaaag gtagatccaa aaaagaagag aaaggtataa
     3541 ctcgagtggc attgaattga attgaaatcg atagatcaat ttttttcttt tctctttccc
     3601 catcctttac gctaaaataa tagtttattt tattttttga atatttttta tttatatacg
     3661 tatatataga ctattattta tcttttaatg attattaaga tttttattaa aaaaaaattc
     3721 gctcctcttt taatgccttt atgcagtttt tttttcccat tcgatatttc tatgtgctgc
     3781 caaaccagat gtcaacacag ctacaacgaa gcttcgtctt ttcttgaaga aaagaggaag
     3841 ctcgctaaat gggattccac tttccgttcc ctgccagctg atggaaaaag gttagtggaa
     3901 cgatgaagaa taaaaagaga gatccactga ggtgaaattt cagctgacag cgagtttcat
     3961 gatcgtgatg aacaatggta acgagttgtg gctgttgcca gggagggtgg ttctcaccaa
     4021 atcgctactt gggtttgaga ttggtaaaat tgattgtttg gatggcaatc atctatggaa
     4081 tggatattat ataacaaaga agaaataatg aactgattct cttcctcctt cttgtccttt
     4141 cttaattctg ttgtaattac cttcctttgt aatttttttt gtaattattc ttcttaataa
     4201 tccaaacaaa cacacatatt acaataagat ctatggtttc aaagggcgaa gaagacaata
     4261 tggctattat taaggaattc atgagattca aggtccacat ggaaggttct gtcaacggtc
     4321 acgaattcga aattgaaggt gaaggtgaag gtagaccata cgaaggtacc caaaccgcta
     4381 agttgaaggt caccaagggt ggtccattgc cattcgcttg ggatattttg tctccacaat
     4441 tcatgtacgg ttctaaggct tacgtcaagc acccagctga tattccagat tacttgaagt
     4501 tgtctttccc agaaggtttc aagtgggaaa gagtcatgaa cttcgaagat ggtggtgtcg
     4561 tcaccgtcac ccaagattct tctttgcaag atggtgaatt catttacaag gtcaagttga
     4621 gaggtaccaa cttcccatct gatggtccag tcatgcaaaa gaagaccatg ggttgggaag
     4681 cttcttctga aagaatgtac ccagaagatg gtgctttgaa gggtgaaatt aagcaaagat
     4741 tgaagttgaa ggatggtggt cactacgatg ctgaagtcaa gaccacctac aaggctaaga
     4801 agccagtcca attgccaggt gcttacaacg tcaacattaa gttggatatt acctctcaca
     4861 acgaagatta caccattgtc gaacaatacg aaagagctga aggtagacac tctaccggtg
     4921 gtatggatga attgtacaag taaatcctaa ctcgagataa agcaatcttg atgaggataa
     4981 tgattttttt ttgaatatac ataaatacta ccgtttttct gctagatttt gtgatgacgt
     5041 aaataagtac atattacttt ttaagccaag acaagattaa gcattaactt tacccttttc
     5101 tttctaagtt tcaatattag ttatcactgt ttaaaagtta tggcgagaac gtcggcggtt
     5161 aaaatatatt accctgaacg gctgagcagt tacagagatg ttacgaacca ctagtgcact
     5221 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     5281 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     5341 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     5401 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     5461 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     5521 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     5581 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     5641 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     5701 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     5761 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     5821 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     5881 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     5941 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     6001 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     6061 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     6121 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     6181 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     6241 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag tatcacgtgc
     6301 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     6361 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     6421 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     6481 attgtttcat cttgtctgtg tagaaaacca cacacgaaaa tcctgtgatt ttacatttta
     6541 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     6601 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     6661 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     6721 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     6781 tacaggcaag cgatccgtc
//
