LOCUS       sequence-258369-        7335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-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 7335)
  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 7335)
  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_096
FEATURES             Location/Qualifiers
     source          1..7335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1290..1310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2250..2270
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2274..2294
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4163..4966
                     /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(5608..6423)
                     /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(5608..6423)
                     /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     6331..6350
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6415..6434
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6648..7236
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7137..7156
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcca atgctaaatt cgagtgaaac acaggaagat cagaaaatcc tcatttcatc
       61 catattaaca ataatttcaa atgtttattt gcattatttg aaactaggca agacaagcaa
      121 cgaaacgttt ttgaaaattt tgagtatttt caataaattt gtagaggact cagatattga
      181 aaaaaagcta cagcaattaa tacttgataa gaagagtatt gagaagggca acggttcatc
      241 atctcatgga tctgcacatg aacaaacacc agagtcaaac gacgttgaaa ttgaggctac
      301 tgcgccaatt gatgacaata cagacgatga taacaaaccg aagttatctg atgtagaaaa
      361 ggattaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt ctatatatgt
      421 taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc atcaaagaag
      481 gttaatgtgg ctgtggtttc agggtccata aagcccacat ggataacatt acccctgaat
      541 tcgcatctag actgaactgg ccgataattg cagacgaacg aagaggatgt ccaatatttt
      601 ttttaaggaa taaggatact tcaagactag attcccccct gcattcccat cagaaccgta
      661 aaccttggcg ctttccttgg gaagtattca agaagtgcct tgtccggttt ctgtggctca
      721 caaaccagcg cgcccgatat ggctttcttt tcacttatga atgtaccagt acgggacaat
      781 tagaacgctc ctgtaacaat ctctttgcaa atgtggggtt acattctaac catgtcacac
      841 tgctgacgaa attcaaagta aaaaaaaatg ggaccacgtc ttgagaacga tagattttct
      901 ttattttaca ttgaacagtc gttgtctcag cgcgctttat gttttcattc atacttcata
      961 ttataaaata acaaaagaag aatttcatat tcacgcccaa gaaatcaggc tgctttccaa
     1021 atgcaattga cacttcatta gccatcacac aaaactcttt cttgctggag cttcttttaa
     1081 aaaagacctc agtacaccaa acacgttacc cgacctcgtt attttacgac aactatgata
     1141 aaattctgaa gaaaaaataa aaaaattttc atacttcttg cttttattta aaccattgaa
     1201 tgatttcttt tgaacaaaac tacctgtttc accaaaggaa atagaaagaa aaaatcaatt
     1261 agaagaaaac aaaaaacaaa agatctatgc caaagaagaa gagaaaggtt ggttctcgtt
     1321 tcaacaagct cgacctcaat cttctggtcg ccctggatgc actgctcacg gagatgagca
     1381 tcagccgcgc cgccgaaaag atccatctga gccagtcggc catgagcaat gccctggcgc
     1441 ggctgcgcga gtatttcgat gatgaattgc tgatccaggt gggccggcgc atggagccca
     1501 cgccgcgcgc cgaggtgctc aaggatgcgg tgcatgatgt gctgcggcgt atcgatggct
     1561 ccatcgcggc gctgccggcc ttcgtgccgg ccgagtccac gcgcgagttt cgcatctcgg
     1621 tttcggactt tacgctctcc gtcctcatcc cccgggtgct ggcgcgcgcg cacgccgagg
     1681 gcaagcacat ccgctttgcc ctgatgccgc aggtgcaaga cccgacccgc tcgctggatc
     1741 gggccgaggt ggacctgctg gtcttgccgc aggaattctg cacgcccgat catcctgccg
     1801 aagaggtctt ccgcgaacgg catgtctgcg tggtctggcg cgacagtgcg ctggcgcaag
     1861 gcgagctgac gctggaacgc tacatggcct caggccatgt ggtgatggtg ccgcctgggg
     1921 ccaatgcgtc gtcggtggag gcgtggatgg ccaggaagct gggctttgcg cgccgggtgg
     1981 aagtgaccag cttcagcttc gcttctgcgc tggcgctggt acaggggacg gaccgcatcg
     2041 ccacggtgca tgcccggctg gcgcagctgc tggctccgca atggccggtg gtgatcaagg
     2101 agagtccgct gtcgctgggc gagatgcggc agatgatgca gtggcatcgc taccgcagca
     2161 atgatcctgg catccagtgg ctgcgtcggg tgtttctgga gagtgcgcag gagatggatg
     2221 cggcgctgcc aggcatctgc ggatccgatc caaaaaagaa gagaaaggta gatccaaaaa
     2281 agaagagaaa ggtataactc gagtggcatt gaattgaatt gaaatcgata gatcaatttt
     2341 tttcttttct ctttccccat cctttacgct aaaataatag tttattttat tttttgaata
     2401 ttttttattt atatacgtat atatagacta ttatttatct tttaatgatt attaagattt
     2461 ttattaaaaa aaaattcgct cctcttttaa tgcctttatg cagttttttt ttcccattcg
     2521 atatttctat gtgctgccaa accagatgtc aacacagcta caacgaagct tcgtcttttc
     2581 ttgaagaaaa gaggaagctc gctaaatggg attccacttt ccgttccctg ccagctgatg
     2641 gaaaaaggtt agtggaacga tgaagaataa aaagagagat ccactgaggt gaaatttcag
     2701 ctgacagcga gtttcatgat cgtgatgaac aatggtaacg agttgtggct gttgccaggg
     2761 agggtggttc tcaccaaatc gctacttggg tttgagattg gtaaaattga ttgtttggat
     2821 ggcaatcatc tatggaatgg atattatata acaaagaaga aataatgaac tgattctctt
     2881 cctccttctt gtcctttctt aattctgttg taattacctt cctttgtaat tttttttgta
     2941 attattcttc ttaataatcc aaacaaacac acatattaca ataagatcta tggtttcaaa
     3001 gggcgaagaa gacaatatgg ctattattaa ggaattcatg agattcaagg tccacatgga
     3061 aggttctgtc aacggtcacg aattcgaaat tgaaggtgaa ggtgaaggta gaccatacga
     3121 aggtacccaa accgctaagt tgaaggtcac caagggtggt ccattgccat tcgcttggga
     3181 tattttgtct ccacaattca tgtacggttc taaggcttac gtcaagcacc cagctgatat
     3241 tccagattac ttgaagttgt ctttcccaga aggtttcaag tgggaaagag tcatgaactt
     3301 cgaagatggt ggtgtcgtca ccgtcaccca agattcttct ttgcaagatg gtgaattcat
     3361 ttacaaggtc aagttgagag gtaccaactt cccatctgat ggtccagtca tgcaaaagaa
     3421 gaccatgggt tgggaagctt cttctgaaag aatgtaccca gaagatggtg ctttgaaggg
     3481 tgaaattaag caaagattga agttgaagga tggtggtcac tacgatgctg aagtcaagac
     3541 cacctacaag gctaagaagc cagtccaatt gccaggtgct tacaacgtca acattaagtt
     3601 ggatattacc tctcacaacg aagattacac cattgtcgaa caatacgaaa gagctgaagg
     3661 tagacactct accggtggta tggatgaatt gtacaagtaa atcctaactc gagataaagc
     3721 aatcttgatg aggataatga tttttttttg aatatacata aatactaccg tttttctgct
     3781 agattttgtg atgacgtaaa taagtacata ttacttttta agccaagaca agattaagca
     3841 ttaactttac ccttttcttt ctaagtttca atattagtta tcactgttta aaagttatgg
     3901 cgagaacgtc ggcggttaaa atatattacc ctgaacggct gagcagttac agagatgtta
     3961 cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc
     4021 cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg
     4081 tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac
     4141 gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca
     4201 tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg
     4261 tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc
     4321 caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac
     4381 agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa
     4441 atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc
     4501 agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt
     4561 gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt
     4621 gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa
     4681 gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga
     4741 aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt
     4801 gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt
     4861 tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
     4921 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
     4981 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
     5041 acccgagtag agcacttgaa tccactgccc cgggaatctc ggtcgtaatg atttctataa
     5101 tgacgaaaaa aaaaaaattg gaaagaaaaa gcttcatggc ctttataaaa aggaactatc
     5161 caatacctcg ccagaaccaa gtaacagtat tttacggggc acaaatcaag aacaataaga
     5221 caggactgta aagatggacg cattgaactc caaagaacaa caagagttcc aaaaagtagt
     5281 ggaacaaaag caaatgaagg atttcatgcg tttgtactct aatctggtag aaagatgttt
     5341 cacagactgt gtcaatgact tcacaacatc aaagctaacc aataaggaac aaacatgcat
     5401 catgaagtgc tcagaaaagt tcttgaagca tagcgaacgt gtagggcagc gtttccaaga
     5461 acaaaacgct gccttgggac aaggcttggg ccgataaggt gtactggcgt atatatatct
     5521 aattatgtat ctctggtgta gcccattttt agcatgtaaa tataaagacc gagcggccgc
     5581 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     5641 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     5701 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     5761 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     5821 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     5881 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     5941 aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt
     6001 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     6061 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     6121 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     6181 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     6241 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     6301 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     6361 agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct
     6421 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     6481 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat
     6541 aaatcgaact tttgctgagt tgaaggatca gtcatgacca aaatccctta acgtgagttt
     6601 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6661 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6721 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6781 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6841 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6901 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6961 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7021 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7081 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7141 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7201 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7261 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7321 tctgtggata accgt
//
