LOCUS       sequence-266872-        7346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pTDH3_ 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 7346)
  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 7346)
  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_007_026_096
FEATURES             Location/Qualifiers
     source          1..7346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             243..263
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             267..287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2156..2959
                     /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(3601..4416)
                     /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(3601..4416)
                     /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     4324..4343
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4408..4427
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4641..5229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5130..5149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5922..6588
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     6458..6481
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             6598..6618
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cgccacggtg catgcccggc tggcgcagct gctggctccg caatggccgg tggtgatcaa
       61 ggagagtccg ctgtcgctgg gcgagatgcg gcagatgatg cagtggcatc gctaccgcag
      121 caatgatcct ggcatccagt ggctgcgtcg ggtgtttctg gagagtgcgc aggagatgga
      181 tgcggcgctg ccaggcatct gcggatcctg agaccaggct gccaggcatc tgcggatccg
      241 atccaaaaaa gaagagaaag gtagatccaa aaaagaagag aaaggtataa ctcgagtggc
      301 attgaattga attgaaatcg atagatcaat ttttttcttt tctctttccc catcctttac
      361 gctaaaataa tagtttattt tattttttga atatttttta tttatatacg tatatataga
      421 ctattattta tcttttaatg attattaaga tttttattaa aaaaaaattc gctcctcttt
      481 taatgccttt atgcagtttt tttttcccat tcgatatttc tatgtgctgc caaaccagat
      541 gtcaacacag ctacaacgaa gcttcgtctt ttcttgaaga aaagaggaag ctcgctaaat
      601 gggattccac tttccgttcc ctgccagctg atggaaaaag gttagtggaa cgatgaagaa
      661 taaaaagaga gatccactga ggtgaaattt cagctgacag cgagtttcat gatcgtgatg
      721 aacaatggta acgagttgtg gctgttgcca gggagggtgg ttctcaccaa atcgctactt
      781 gggtttgaga ttggtaaaat tgattgtttg gatggcaatc atctatggaa tggatattat
      841 ataacaaaga agaaataatg aactgattct cttcctcctt cttgtccttt cttaattctg
      901 ttgtaattac cttcctttgt aatttttttt gtaattattc ttcttaataa tccaaacaaa
      961 cacacatatt acaataagat ctatggtttc aaagggcgaa gaagacaata tggctattat
     1021 taaggaattc atgagattca aggtccacat ggaaggttct gtcaacggtc acgaattcga
     1081 aattgaaggt gaaggtgaag gtagaccata cgaaggtacc caaaccgcta agttgaaggt
     1141 caccaagggt ggtccattgc cattcgcttg ggatattttg tctccacaat tcatgtacgg
     1201 ttctaaggct tacgtcaagc acccagctga tattccagat tacttgaagt tgtctttccc
     1261 agaaggtttc aagtgggaaa gagtcatgaa cttcgaagat ggtggtgtcg tcaccgtcac
     1321 ccaagattct tctttgcaag atggtgaatt catttacaag gtcaagttga gaggtaccaa
     1381 cttcccatct gatggtccag tcatgcaaaa gaagaccatg ggttgggaag cttcttctga
     1441 aagaatgtac ccagaagatg gtgctttgaa gggtgaaatt aagcaaagat tgaagttgaa
     1501 ggatggtggt cactacgatg ctgaagtcaa gaccacctac aaggctaaga agccagtcca
     1561 attgccaggt gcttacaacg tcaacattaa gttggatatt acctctcaca acgaagatta
     1621 caccattgtc gaacaatacg aaagagctga aggtagacac tctaccggtg gtatggatga
     1681 attgtacaag taaatcctaa ctcgagataa agcaatcttg atgaggataa tgattttttt
     1741 ttgaatatac ataaatacta ccgtttttct gctagatttt gtgatgacgt aaataagtac
     1801 atattacttt ttaagccaag acaagattaa gcattaactt tacccttttc tttctaagtt
     1861 tcaatattag ttatcactgt ttaaaagtta tggcgagaac gtcggcggtt aaaatatatt
     1921 accctgaacg gctgagcagt tacagagatg ttacgaacca ctagtgcact gcagtacacg
     1981 gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
     2041 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     2101 agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc
     2161 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     2221 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     2281 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     2341 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     2401 caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa
     2461 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     2521 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa
     2581 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg
     2641 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
     2701 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     2761 cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga
     2821 tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     2881 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     2941 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     3001 aattagagct tcaatttaat tatatcagtt attacccgag tagagcactt gaatccactg
     3061 ccccgggaat ctcggtcgta atgatttcta taatgacgaa aaaaaaaaaa ttggaaagaa
     3121 aaagcttcat ggcctttata aaaaggaact atccaatacc tcgccagaac caagtaacag
     3181 tattttacgg ggcacaaatc aagaacaata agacaggact gtaaagatgg acgcattgaa
     3241 ctccaaagaa caacaagagt tccaaaaagt agtggaacaa aagcaaatga aggatttcat
     3301 gcgtttgtac tctaatctgg tagaaagatg tttcacagac tgtgtcaatg acttcacaac
     3361 atcaaagcta accaataagg aacaaacatg catcatgaag tgctcagaaa agttcttgaa
     3421 gcatagcgaa cgtgtagggc agcgtttcca agaacaaaac gctgccttgg gacaaggctt
     3481 gggccgataa ggtgtactgg cgtatatata tctaattatg tatctctggt gtagcccatt
     3541 tttagcatgt aaatataaag accgagcggc cgcgtgttac aaccaattaa ccaattctga
     3601 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     3661 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     3721 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     3781 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     3841 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     3901 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     3961 cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     4021 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     4081 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     4141 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     4201 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     4261 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     4321 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     4381 cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     4441 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     4501 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
     4561 tcagtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4621 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4681 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4741 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4801 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4861 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4921 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4981 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5041 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5101 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5161 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5221 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5281 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgccaat
     5341 gctaaattcg agtgaaacac aggaagatca gaaaatcctc atttcatcca tattaacaat
     5401 aatttcaaat gtttatttgc attatttgaa actaggcaag acaagcaacg aaacgttttt
     5461 gaaaattttg agtattttca ataaatttgt agaggactca gatattgaaa aaaagctaca
     5521 gcaattaata cttgataaga agagtattga gaagggcaac ggttcatcat ctcatggatc
     5581 tgcacatgaa caaacaccag agtcaaacga cgttgaaatt gaggctactg cgccaattga
     5641 tgacaataca gacgatgata acaaaccgaa gttatctgat gtagaaaagg attaaagatg
     5701 ctaagagata gtgatgatat ttcataaata atgtaattct atatatgtta attacctttt
     5761 ttgcgaggca tatttatggt gaaggataag ttttgaccat caaagaaggt taatgtggct
     5821 gtggtttcag ggtccataaa gcccacatgg ataacattac ccctgaattc gcatctagac
     5881 tgaactggcc gataattgca gacgaacgca gttcgagttt atcattatca atactgccat
     5941 ttcaaagaat acgtaaataa ttaatagtag tgattttcct aactttattt agtcaaaaaa
     6001 ttagcctttt aattctgctg taacccgtac atgcccaaaa tagggggcgg gttacacaga
     6061 atatataaca tcgtaggtgt ctgggtgaac agtttattcc tggcatccac taaatataat
     6121 ggagcccgct ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt
     6181 tttcttcacc aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg
     6241 gcacaaacag gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa
     6301 atgatgacac aaggcaattg acccacgcat gtatctatct cattttctta caccttctat
     6361 taccttctgc tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg
     6421 aaattattcc cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct
     6481 gtaaatctat ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt
     6541 taaaacacca agaacttagt ttcgaataaa cacacataaa caaacaaaag atctatgcca
     6601 aagaagaaga gaaaggttgg ttctcgtttc aacaagctcg acctcaatct tctggtcgcc
     6661 ctggatgcac tgctcacgga gatgagcatc agccgcgccg ccgaaaagat ccatctgagc
     6721 cagtcggcca tgagcaatgc cctggcgcgg ctgcgcgagt atttcgatga tgaattgctg
     6781 atccaggtgg gccggcgcat ggagcccacg ccgcgcgccg aggtgctcaa ggatgcggtg
     6841 catgatgtgc tgcggcgtat cgatggctcc atcgcggcgc tgccggcctt cgtgccggcc
     6901 gagtccacgc gcgagtttcg catctcggtt tcggacttta cgctctccgt cctcatcccc
     6961 cgggtgctgg cgcgcgcgca cgccgagggc aagcacatcc gctttgccct gatgccgcag
     7021 gtgcaagacc cgacccgctc gctggatcgg gccgaggtgg acctgctggt cttgccgcag
     7081 gaattctgca cgcccgatca tcctgccgaa gaggtcttcc gcgaacggca tgtctgcgtg
     7141 gtctggcgcg acagtgcgct ggcgcaaggc gagctgacgc tggaacgcta catggcctca
     7201 ggccatgtgg tgatggtgcc gcctggggcc aatgcgtcgt cggtggaggc gtggatggcc
     7261 aggaagctgg gctttgcgcg ccgggtggaa gtgaccagct tcagcttcgc ttctgcgctg
     7321 gcgctggtac aggggacgga ccgcat
//
