LOCUS       sequence-266974-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-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 7284)
  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 7284)
  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_096
FEATURES             Location/Qualifiers
     source          1..7284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             785..805
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3607..4410
                     /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"
     CDS             complement(5052..5867)
                     /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(5052..5867)
                     /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     5775..5794
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5859..5878
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6092..6680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6581..6600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccataaagcc cacatggata acattacccc tgaattcgca tctagactga actggccgat
       61 aattgcagac gaacgaagag gatgtccaat atttttttta aggaataagg atacttcaag
      121 actagattcc cccctgcatt cccatcagaa ccgtaaacct tggcgctttc cttgggaagt
      181 attcaagaag tgccttgtcc ggtttctgtg gctcacaaac cagcgcgccc gatatggctt
      241 tcttttcact tatgaatgta ccagtacggg acaattagaa cgctcctgta acaatctctt
      301 tgcaaatgtg gggttacatt ctaaccatgt cacactgctg acgaaattca aagtaaaaaa
      361 aaatgggacc acgtcttgag aacgatagat tttctttatt ttacattgaa cagtcgttgt
      421 ctcagcgcgc tttatgtttt cattcatact tcatattata aaataacaaa agaagaattt
      481 catattcacg cccaagaaat caggctgctt tccaaatgca attgacactt cattagccat
      541 cacacaaaac tctttcttgc tggagcttct tttaaaaaag acctcagtac accaaacacg
      601 ttacccgacc tcgttatttt acgacaacta tgataaaatt ctgaagaaaa aataaaaaaa
      661 ttttcatact tcttgctttt atttaaacca ttgaatgatt tcttttgaac aaaactacct
      721 gtttcaccaa aggaaataga aagaaaaaat caattagaag aaaacaaaaa acaaaagatc
      781 tatgccaaag aagaagagaa aggttggttc tcgtttcaac aagctcgacc tcaatcttct
      841 ggtcgccctg gatgcactgc tcacggagat gagcatcagc cgcgccgccg aaaagatcca
      901 tctgagccag tcggccatga gcaatgccct ggcgcggctg cgcgagtatt tcgatgatga
      961 attgctgatc caggtgggcc ggcgcatgga gcccacgccg cgcgccgagg tgctcaagga
     1021 tgcggtgcat gatgtgctgc ggcgtatcga tggctccatc gcggcgctgc cggccttcgt
     1081 gccggccgag tccacgcgcg agtttcgcat ctcggtttcg gactttacgc tctccgtcct
     1141 catcccccgg gtgctggcgc gcgcgcacgc cgagggcaag cacatccgct ttgccctgat
     1201 gccgcaggtg caagacccga cccgctcgct ggatcgggcc gaggtggacc tgctggtctt
     1261 gccgcaggaa ttctgcacgc ccgatcatcc tgccgaagag gtcttccgcg aacggcatgt
     1321 ctgcgtggtc tggcgcgaca gtgcgctggc gcaaggcgag ctgacgctgg aacgctacat
     1381 ggcctcaggc catgtggtga tggtgccgcc tggggccaat gcgtcgtcgg tggaggcgtg
     1441 gatggccagg aagctgggct ttgcgcgccg ggtggaagtg accagcttca gcttcgcttc
     1501 tgcgctggcg ctggtacagg ggacggaccg catcgccacg gtgcatgccc ggctggcgca
     1561 gctgctggct ccgcaatggc cggtggtgat caaggagagt ccgctgtcgc tgggcgagat
     1621 gcggcagatg atgcagtggc atcgctaccg cagcaatgat cctggcatcc agtggctgcg
     1681 tcgggtgttt ctggagagtg cgcaggagat ggatgcggcg ctgccaggca tctgctgaat
     1741 cctaactcga gattgaattg aattgaaatc gatagatcaa tttttttctt ttctctttcc
     1801 ccatccttta cgctaaaata atagtttatt ttattttttg aatatttttt atttatatac
     1861 gtatatatag actattattt atcttttaat gattattaag atttttatta aaaaaaaatt
     1921 cgctcctctt ttaatgcctt tatgcagttt ttttttccca ttcgatattt ctatgtgctg
     1981 ccaaaccaga tgtcaacaca gctacaacga agcttcgtct tttcttgaag aaaagaggaa
     2041 gctcgctaaa tgggattcca ctttccgttc cctgccagct gatggaaaaa ggttagtgga
     2101 acgatgaaga ataaaaagag agatccactg aggtgaaatt tcagctgaca gcgagtttca
     2161 tgatcgtgat gaacaatggt aacgagttgt ggctgttgcc agggagggtg gttctcacca
     2221 aatcgctact tgggtttgag attggtaaaa ttgattgttt ggatggcaat catctatgga
     2281 atggatatta tataacaaag aagaaataat gaactgattc tcttcctcct tcttgtcctt
     2341 tcttaattct gttgtaatta ccttcctttg taattttttt tgtaattatt cttcttaata
     2401 atccaaacaa acacacatat tacaataaga tctatggttt caaagggcga agaagacaat
     2461 atggctatta ttaaggaatt catgagattc aaggtccaca tggaaggttc tgtcaacggt
     2521 cacgaattcg aaattgaagg tgaaggtgaa ggtagaccat acgaaggtac ccaaaccgct
     2581 aagttgaagg tcaccaaggg tggtccattg ccattcgctt gggatatttt gtctccacaa
     2641 ttcatgtacg gttctaaggc ttacgtcaag cacccagctg atattccaga ttacttgaag
     2701 ttgtctttcc cagaaggttt caagtgggaa agagtcatga acttcgaaga tggtggtgtc
     2761 gtcaccgtca cccaagattc ttctttgcaa gatggtgaat tcatttacaa ggtcaagttg
     2821 agaggtacca acttcccatc tgatggtcca gtcatgcaaa agaagaccat gggttgggaa
     2881 gcttcttctg aaagaatgta cccagaagat ggtgctttga agggtgaaat taagcaaaga
     2941 ttgaagttga aggatggtgg tcactacgat gctgaagtca agaccaccta caaggctaag
     3001 aagccagtcc aattgccagg tgcttacaac gtcaacatta agttggatat tacctctcac
     3061 aacgaagatt acaccattgt cgaacaatac gaaagagctg aaggtagaca ctctaccggt
     3121 ggtatggatg aattgtacaa gtaaatccta actcgagata aagcaatctt gatgaggata
     3181 atgatttttt tttgaatata cataaatact accgtttttc tgctagattt tgtgatgacg
     3241 taaataagta catattactt tttaagccaa gacaagatta agcattaact ttaccctttt
     3301 ctttctaagt ttcaatatta gttatcactg tttaaaagtt atggcgagaa cgtcggcggt
     3361 taaaatatat taccctgaac ggctgagcag ttacagagat gttacgaacc actagtgcac
     3421 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     3481 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     3541 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     3601 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     3661 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     3721 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     3781 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     3841 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     3901 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     3961 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     4021 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     4081 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     4141 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     4201 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     4261 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     4321 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     4381 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     4441 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtagagcact
     4501 tgaatccact gccccgggaa tctcggtcgt aatgatttct ataatgacga aaaaaaaaaa
     4561 attggaaaga aaaagcttca tggcctttat aaaaaggaac tatccaatac ctcgccagaa
     4621 ccaagtaaca gtattttacg gggcacaaat caagaacaat aagacaggac tgtaaagatg
     4681 gacgcattga actccaaaga acaacaagag ttccaaaaag tagtggaaca aaagcaaatg
     4741 aaggatttca tgcgtttgta ctctaatctg gtagaaagat gtttcacaga ctgtgtcaat
     4801 gacttcacaa catcaaagct aaccaataag gaacaaacat gcatcatgaa gtgctcagaa
     4861 aagttcttga agcatagcga acgtgtaggg cagcgtttcc aagaacaaaa cgctgccttg
     4921 ggacaaggct tgggccgata aggtgtactg gcgtatatat atctaattat gtatctctgg
     4981 tgtagcccat ttttagcatg taaatataaa gaccgagcgg ccgcgtgtta caaccaatta
     5041 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     5101 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     5161 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     5221 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     5281 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     5341 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     5401 attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa
     5461 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     5521 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     5581 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     5641 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     5701 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     5761 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     5821 tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     5881 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     5941 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     6001 gagttgaagg atcagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6061 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6121 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6181 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6601 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6661 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6721 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
     6781 cggccgccaa tgctaaattc gagtgaaaca caggaagatc agaaaatcct catttcatcc
     6841 atattaacaa taatttcaaa tgtttatttg cattatttga aactaggcaa gacaagcaac
     6901 gaaacgtttt tgaaaatttt gagtattttc aataaatttg tagaggactc agatattgaa
     6961 aaaaagctac agcaattaat acttgataag aagagtattg agaagggcaa cggttcatca
     7021 tctcatggat ctgcacatga acaaacacca gagtcaaacg acgttgaaat tgaggctact
     7081 gcgccaattg atgacaatac agacgatgat aacaaaccga agttatctga tgtagaaaag
     7141 gattaaagat gctaagagat agtgatgata tttcataaat aatgtaattc tatatatgtt
     7201 aattaccttt tttgcgaggc atatttatgg tgaaggataa gttttgacca tcaaagaagg
     7261 ttaatgtggc tgtggtttca gggt
//
