LOCUS       sequence-266910-        6772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pREV1_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 6772)
  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 6772)
  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_006_026_096
FEATURES             Location/Qualifiers
     source          1..6772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..24
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1893..2696
                     /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    2779..3282
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(3326..4141)
                     /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(3326..4141)
                     /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     4049..4068
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4133..4152
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4366..4954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4855..4874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6752..6772
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gatccaaaaa agaagagaaa ggtataactc gagtggcatt gaattgaatt gaaatcgata
       61 gatcaatttt tttcttttct ctttccccat cctttacgct aaaataatag tttattttat
      121 tttttgaata ttttttattt atatacgtat atatagacta ttatttatct tttaatgatt
      181 attaagattt ttattaaaaa aaaattcgct cctcttttaa tgcctttatg cagttttttt
      241 ttcccattcg atatttctat gtgctgccaa accagatgtc aacacagcta caacgaagct
      301 tcgtcttttc ttgaagaaaa gaggaagctc gctaaatggg attccacttt ccgttccctg
      361 ccagctgatg gaaaaaggtt agtggaacga tgaagaataa aaagagagat ccactgaggt
      421 gaaatttcag ctgacagcga gtttcatgat cgtgatgaac aatggtaacg agttgtggct
      481 gttgccaggg agggtggttc tcaccaaatc gctacttggg tttgagattg gtaaaattga
      541 ttgtttggat ggcaatcatc tatggaatgg atattatata acaaagaaga aataatgaac
      601 tgattctctt cctccttctt gtcctttctt aattctgttg taattacctt cctttgtaat
      661 tttttttgta attattcttc ttaataatcc aaacaaacac acatattaca ataagatcta
      721 tggtttcaaa gggcgaagaa gacaatatgg ctattattaa ggaattcatg agattcaagg
      781 tccacatgga aggttctgtc aacggtcacg aattcgaaat tgaaggtgaa ggtgaaggta
      841 gaccatacga aggtacccaa accgctaagt tgaaggtcac caagggtggt ccattgccat
      901 tcgcttggga tattttgtct ccacaattca tgtacggttc taaggcttac gtcaagcacc
      961 cagctgatat tccagattac ttgaagttgt ctttcccaga aggtttcaag tgggaaagag
     1021 tcatgaactt cgaagatggt ggtgtcgtca ccgtcaccca agattcttct ttgcaagatg
     1081 gtgaattcat ttacaaggtc aagttgagag gtaccaactt cccatctgat ggtccagtca
     1141 tgcaaaagaa gaccatgggt tgggaagctt cttctgaaag aatgtaccca gaagatggtg
     1201 ctttgaaggg tgaaattaag caaagattga agttgaagga tggtggtcac tacgatgctg
     1261 aagtcaagac cacctacaag gctaagaagc cagtccaatt gccaggtgct tacaacgtca
     1321 acattaagtt ggatattacc tctcacaacg aagattacac cattgtcgaa caatacgaaa
     1381 gagctgaagg tagacactct accggtggta tggatgaatt gtacaagtaa atcctaactc
     1441 gagataaagc aatcttgatg aggataatga tttttttttg aatatacata aatactaccg
     1501 tttttctgct agattttgtg atgacgtaaa taagtacata ttacttttta agccaagaca
     1561 agattaagca ttaactttac ccttttcttt ctaagtttca atattagtta tcactgttta
     1621 aaagttatgg cgagaacgtc ggcggttaaa atatattacc ctgaacggct gagcagttac
     1681 agagatgtta cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     1741 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     1801 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     1861 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     1921 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     1981 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     2041 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     2101 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     2161 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     2221 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     2281 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     2341 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     2401 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     2461 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     2521 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     2581 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     2641 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     2701 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     2761 atcagttatt acccgagtat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     2821 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     2881 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     2941 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aaaaccacac
     3001 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     3061 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     3121 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     3181 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     3241 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcccga gcggccgcgt
     3301 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     3361 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     3421 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     3481 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     3541 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     3601 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     3661 ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa
     3721 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     3781 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     3841 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     3901 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     3961 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     4021 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     4081 catccatgtt ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca
     4141 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     4201 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     4261 atcgaacttt tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagttttc
     4321 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4381 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4441 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4501 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4561 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4621 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4681 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4741 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4801 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4861 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4921 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4981 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5041 tgtggataac cgtgcggccg cccctgaatt cgcatctaga ctgaactggc cgataattgc
     5101 agacgaacgg tgttgttatc cgatacaacc ggatattttt cttttaatga gtctaaaccg
     5161 tgatagcttc aggttaatac aatcaaaaaa agctcaaata ttcttttaat gccgcgttca
     5221 cagattccaa ttgaatacaa ctaggtagtt cattatatga agcctttgct actatttttc
     5281 actatagtct gccttcacct taatgcagac atccacatat tttaatcact ttaaaataaa
     5341 aaggaagata tattagaagc tatgatccaa tctgtaagcc agattaaaat tcacgaactc
     5401 ttctttcatt tgaattgaat gctttgagtt ggggtagatt atcgcaaatt actcatcaca
     5461 tttattgact acgaacttgc tgatgtcctt tttttattta tatttttctt cagtgaagcg
     5521 attttttttt tacacagacc aagacggaaa aaagtagcta aggaagaaaa caaaatcatg
     5581 aaaaaaatgt gaagtgatca tgcacatcgc atcaacttaa acattggctt agagatatat
     5641 agagttagag tttacggcaa cctttaagca ccaatacctt ttggcatagt ctaaagacct
     5701 ggttcttaat tttaaacaaa tttaactaaa gatttcccta tcaaagaagt aacgagttga
     5761 cagattttct caaaataaat cgatactgca tttctaggca tatccagcga gatctatgcg
     5821 tttcaacaag ctcgacctca atcttctggt cgccctggat gcactgctca cggagatgag
     5881 catcagccgc gccgccgaaa agatccatct gagccagtcg gccatgagca atgccctggc
     5941 gcggctgcgc gagtatttcg atgatgaatt gctgatccag gtgggccggc gcatggagcc
     6001 cacgccgcgc gccgaggtgc tcaaggatgc ggtgcatgat gtgctgcggc gtatcgatgg
     6061 ctccatcgcg gcgctgccgg ccttcgtgcc ggccgagtcc acgcgcgagt ttcgcatctc
     6121 ggtttcggac tttacgctct ccgtcctcat cccccgggtg ctggcgcgcg cgcacgccga
     6181 gggcaagcac atccgctttg ccctgatgcc gcaggtgcaa gacccgaccc gctcgctgga
     6241 tcgggccgag gtggacctgc tggtcttgcc gcaggaattc tgcacgcccg atcatcctgc
     6301 cgaagaggtc ttccgcgaac ggcatgtctg cgtggtctgg cgcgacagtg cgctggcgca
     6361 aggcgagctg acgctggaac gctacatggc ctcaggccat gtggtgatgg tgccgcctgg
     6421 ggccaatgcg tcgtcggtgg aggcgtggat ggccaggaag ctgggctttg cgcgccgggt
     6481 ggaagtgacc agcttcagct tcgcttctgc gctggcgctg gtacagggga cggaccgcat
     6541 cgccacggtg catgcccggc tggcgcagct gctggctccg caatggccgg tggtgatcaa
     6601 ggagagtccg ctgtcgctgg gcgagatgcg gcagatgatg cagtggcatc gctaccgcag
     6661 caatgatcct ggcatccagt ggctgcgtcg ggtgtttctg gagagtgcgc aggagatgga
     6721 tgcggcgctg ccaggcatct gcggatccga tccaaaaaag aagagaaagg ta
//
