LOCUS       sequence-258365-        7308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pRPL18B_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 7308)
  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 7308)
  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_005_026_096
FEATURES             Location/Qualifiers
     source          1..7308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2223..2243
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2247..2267
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4136..4939
                     /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(5581..6396)
                     /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(5581..6396)
                     /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     6304..6323
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6388..6407
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6621..7209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7110..7129
                     /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 gtttcaacaa gctcgacctc aatcttctgg
     1321 tcgccctgga tgcactgctc acggagatga gcatcagccg cgccgccgaa aagatccatc
     1381 tgagccagtc ggccatgagc aatgccctgg cgcggctgcg cgagtatttc gatgatgaat
     1441 tgctgatcca ggtgggccgg cgcatggagc ccacgccgcg cgccgaggtg ctcaaggatg
     1501 cggtgcatga tgtgctgcgg cgtatcgatg gctccatcgc ggcgctgccg gccttcgtgc
     1561 cggccgagtc cacgcgcgag tttcgcatct cggtttcgga ctttacgctc tccgtcctca
     1621 tcccccgggt gctggcgcgc gcgcacgccg agggcaagca catccgcttt gccctgatgc
     1681 cgcaggtgca agacccgacc cgctcgctgg atcgggccga ggtggacctg ctggtcttgc
     1741 cgcaggaatt ctgcacgccc gatcatcctg ccgaagaggt cttccgcgaa cggcatgtct
     1801 gcgtggtctg gcgcgacagt gcgctggcgc aaggcgagct gacgctggaa cgctacatgg
     1861 cctcaggcca tgtggtgatg gtgccgcctg gggccaatgc gtcgtcggtg gaggcgtgga
     1921 tggccaggaa gctgggcttt gcgcgccggg tggaagtgac cagcttcagc ttcgcttctg
     1981 cgctggcgct ggtacagggg acggaccgca tcgccacggt gcatgcccgg ctggcgcagc
     2041 tgctggctcc gcaatggccg gtggtgatca aggagagtcc gctgtcgctg ggcgagatgc
     2101 ggcagatgat gcagtggcat cgctaccgca gcaatgatcc tggcatccag tggctgcgtc
     2161 gggtgtttct ggagagtgcg caggagatgg atgcggcgct gccaggcatc tgcggatccg
     2221 atccaaaaaa gaagagaaag gtagatccaa aaaagaagag aaaggtataa ctcgagtggc
     2281 attgaattga attgaaatcg atagatcaat ttttttcttt tctctttccc catcctttac
     2341 gctaaaataa tagtttattt tattttttga atatttttta tttatatacg tatatataga
     2401 ctattattta tcttttaatg attattaaga tttttattaa aaaaaaattc gctcctcttt
     2461 taatgccttt atgcagtttt tttttcccat tcgatatttc tatgtgctgc caaaccagat
     2521 gtcaacacag ctacaacgaa gcttcgtctt ttcttgaaga aaagaggaag ctcgctaaat
     2581 gggattccac tttccgttcc ctgccagctg atggaaaaag gttagtggaa cgatgaagaa
     2641 taaaaagaga gatccactga ggtgaaattt cagctgacag cgagtttcat gatcgtgatg
     2701 aacaatggta acgagttgtg gctgttgcca gggagggtgg ttctcaccaa atcgctactt
     2761 gggtttgaga ttggtaaaat tgattgtttg gatggcaatc atctatggaa tggatattat
     2821 ataacaaaga agaaataatg aactgattct cttcctcctt cttgtccttt cttaattctg
     2881 ttgtaattac cttcctttgt aatttttttt gtaattattc ttcttaataa tccaaacaaa
     2941 cacacatatt acaataagat ctatggtttc aaagggcgaa gaagacaata tggctattat
     3001 taaggaattc atgagattca aggtccacat ggaaggttct gtcaacggtc acgaattcga
     3061 aattgaaggt gaaggtgaag gtagaccata cgaaggtacc caaaccgcta agttgaaggt
     3121 caccaagggt ggtccattgc cattcgcttg ggatattttg tctccacaat tcatgtacgg
     3181 ttctaaggct tacgtcaagc acccagctga tattccagat tacttgaagt tgtctttccc
     3241 agaaggtttc aagtgggaaa gagtcatgaa cttcgaagat ggtggtgtcg tcaccgtcac
     3301 ccaagattct tctttgcaag atggtgaatt catttacaag gtcaagttga gaggtaccaa
     3361 cttcccatct gatggtccag tcatgcaaaa gaagaccatg ggttgggaag cttcttctga
     3421 aagaatgtac ccagaagatg gtgctttgaa gggtgaaatt aagcaaagat tgaagttgaa
     3481 ggatggtggt cactacgatg ctgaagtcaa gaccacctac aaggctaaga agccagtcca
     3541 attgccaggt gcttacaacg tcaacattaa gttggatatt acctctcaca acgaagatta
     3601 caccattgtc gaacaatacg aaagagctga aggtagacac tctaccggtg gtatggatga
     3661 attgtacaag taaatcctaa ctcgagataa agcaatcttg atgaggataa tgattttttt
     3721 ttgaatatac ataaatacta ccgtttttct gctagatttt gtgatgacgt aaataagtac
     3781 atattacttt ttaagccaag acaagattaa gcattaactt tacccttttc tttctaagtt
     3841 tcaatattag ttatcactgt ttaaaagtta tggcgagaac gtcggcggtt aaaatatatt
     3901 accctgaacg gctgagcagt tacagagatg ttacgaacca ctagtgcact gcagtacacg
     3961 gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
     4021 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     4081 agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc
     4141 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     4201 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     4261 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     4321 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     4381 caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa
     4441 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     4501 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa
     4561 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg
     4621 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
     4681 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     4741 cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga
     4801 tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     4861 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     4921 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     4981 aattagagct tcaatttaat tatatcagtt attacccgag tagagcactt gaatccactg
     5041 ccccgggaat ctcggtcgta atgatttcta taatgacgaa aaaaaaaaaa ttggaaagaa
     5101 aaagcttcat ggcctttata aaaaggaact atccaatacc tcgccagaac caagtaacag
     5161 tattttacgg ggcacaaatc aagaacaata agacaggact gtaaagatgg acgcattgaa
     5221 ctccaaagaa caacaagagt tccaaaaagt agtggaacaa aagcaaatga aggatttcat
     5281 gcgtttgtac tctaatctgg tagaaagatg tttcacagac tgtgtcaatg acttcacaac
     5341 atcaaagcta accaataagg aacaaacatg catcatgaag tgctcagaaa agttcttgaa
     5401 gcatagcgaa cgtgtagggc agcgtttcca agaacaaaac gctgccttgg gacaaggctt
     5461 gggccgataa ggtgtactgg cgtatatata tctaattatg tatctctggt gtagcccatt
     5521 tttagcatgt aaatataaag accgagcggc cgcgtgttac aaccaattaa ccaattctga
     5581 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     5641 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     5701 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     5761 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     5821 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     5881 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     5941 cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     6001 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     6061 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     6121 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     6181 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     6241 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     6301 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     6361 cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     6421 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     6481 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
     6541 tcagtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6601 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6661 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6721 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6781 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6841 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6901 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6961 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7021 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7081 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7141 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7201 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7261 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgt
//
