LOCUS       sequence-258363-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pREV1_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_003_026_096
FEATURES             Location/Qualifiers
     source          1..7284
                     /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             4112..4915
                     /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(5557..6372)
                     /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(5557..6372)
                     /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     6280..6299
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6364..6383
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6597..7185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7086..7105
                     /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 gtgttgttat ccgatacaac
      601 cggatatttt tcttttaatg agtctaaacc gtgatagctt caggttaata caatcaaaaa
      661 aagctcaaat attcttttaa tgccgcgttc acagattcca attgaataca actaggtagt
      721 tcattatatg aagcctttgc tactattttt cactatagtc tgccttcacc ttaatgcaga
      781 catccacata ttttaatcac tttaaaataa aaaggaagat atattagaag ctatgatcca
      841 atctgtaagc cagattaaaa ttcacgaact cttctttcat ttgaattgaa tgctttgagt
      901 tggggtagat tatcgcaaat tactcatcac atttattgac tacgaacttg ctgatgtcct
      961 ttttttattt atatttttct tcagtgaagc gatttttttt ttacacagac caagacggaa
     1021 aaaagtagct aaggaagaaa acaaaatcat gaaaaaaatg tgaagtgatc atgcacatcg
     1081 catcaactta aacattggct tagagatata tagagttaga gtttacggca acctttaagc
     1141 accaatacct tttggcatag tctaaagacc tggttcttaa ttttaaacaa atttaactaa
     1201 agatttccct atcaaagaag taacgagttg acagattttc tcaaaataaa tcgatactgc
     1261 atttctaggc atatccagcg 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 tgaatcctaa ctcgagattg aattgaattg aaatcgatag
     2281 atcaattttt ttcttttctc tttccccatc ctttacgcta aaataatagt ttattttatt
     2341 ttttgaatat tttttattta tatacgtata tatagactat tatttatctt ttaatgatta
     2401 ttaagatttt tattaaaaaa aaattcgctc ctcttttaat gcctttatgc agtttttttt
     2461 tcccattcga tatttctatg tgctgccaaa ccagatgtca acacagctac aacgaagctt
     2521 cgtcttttct tgaagaaaag aggaagctcg ctaaatggga ttccactttc cgttccctgc
     2581 cagctgatgg aaaaaggtta gtggaacgat gaagaataaa aagagagatc cactgaggtg
     2641 aaatttcagc tgacagcgag tttcatgatc gtgatgaaca atggtaacga gttgtggctg
     2701 ttgccaggga gggtggttct caccaaatcg ctacttgggt ttgagattgg taaaattgat
     2761 tgtttggatg gcaatcatct atggaatgga tattatataa caaagaagaa ataatgaact
     2821 gattctcttc ctccttcttg tcctttctta attctgttgt aattaccttc ctttgtaatt
     2881 ttttttgtaa ttattcttct taataatcca aacaaacaca catattacaa taagatctat
     2941 ggtttcaaag ggcgaagaag acaatatggc tattattaag gaattcatga gattcaaggt
     3001 ccacatggaa ggttctgtca acggtcacga attcgaaatt gaaggtgaag gtgaaggtag
     3061 accatacgaa ggtacccaaa ccgctaagtt gaaggtcacc aagggtggtc cattgccatt
     3121 cgcttgggat attttgtctc cacaattcat gtacggttct aaggcttacg tcaagcaccc
     3181 agctgatatt ccagattact tgaagttgtc tttcccagaa ggtttcaagt gggaaagagt
     3241 catgaacttc gaagatggtg gtgtcgtcac cgtcacccaa gattcttctt tgcaagatgg
     3301 tgaattcatt tacaaggtca agttgagagg taccaacttc ccatctgatg gtccagtcat
     3361 gcaaaagaag accatgggtt gggaagcttc ttctgaaaga atgtacccag aagatggtgc
     3421 tttgaagggt gaaattaagc aaagattgaa gttgaaggat ggtggtcact acgatgctga
     3481 agtcaagacc acctacaagg ctaagaagcc agtccaattg ccaggtgctt acaacgtcaa
     3541 cattaagttg gatattacct ctcacaacga agattacacc attgtcgaac aatacgaaag
     3601 agctgaaggt agacactcta ccggtggtat ggatgaattg tacaagtaaa tcctaactcg
     3661 agataaagca atcttgatga ggataatgat ttttttttga atatacataa atactaccgt
     3721 ttttctgcta gattttgtga tgacgtaaat aagtacatat tactttttaa gccaagacaa
     3781 gattaagcat taactttacc cttttctttc taagtttcaa tattagttat cactgtttaa
     3841 aagttatggc gagaacgtcg gcggttaaaa tatattaccc tgaacggctg agcagttaca
     3901 gagatgttac gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc
     3961 ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat
     4021 acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag
     4081 aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc
     4141 tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac
     4201 aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc
     4261 attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat
     4321 ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga
     4381 ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata
     4441 cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt
     4501 tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt
     4561 agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga
     4621 cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg
     4681 aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg
     4741 agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat
     4801 tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
     4861 ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
     4921 tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
     4981 tcagttatta cccgagtaga gcacttgaat ccactgcccc gggaatctcg gtcgtaatga
     5041 tttctataat gacgaaaaaa aaaaaattgg aaagaaaaag cttcatggcc tttataaaaa
     5101 ggaactatcc aatacctcgc cagaaccaag taacagtatt ttacggggca caaatcaaga
     5161 acaataagac aggactgtaa agatggacgc attgaactcc aaagaacaac aagagttcca
     5221 aaaagtagtg gaacaaaagc aaatgaagga tttcatgcgt ttgtactcta atctggtaga
     5281 aagatgtttc acagactgtg tcaatgactt cacaacatca aagctaacca ataaggaaca
     5341 aacatgcatc atgaagtgct cagaaaagtt cttgaagcat agcgaacgtg tagggcagcg
     5401 tttccaagaa caaaacgctg ccttgggaca aggcttgggc cgataaggtg tactggcgta
     5461 tatatatcta attatgtatc tctggtgtag cccattttta gcatgtaaat ataaagaccg
     5521 agcggccgcg tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     5581 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     5641 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     5701 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     5761 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     5821 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     5881 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc gaaatacgcg
     5941 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     6001 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     6061 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     6121 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     6181 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     6241 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     6301 atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg
     6361 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     6421 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     6481 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag tcatgaccaa aatcccttaa
     6541 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6601 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6661 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6721 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6781 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6841 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6901 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6961 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7021 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7081 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7141 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7201 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7261 tcccctgatt ctgtggataa ccgt
//
