LOCUS       sequence-266888-        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             2802..3605
                     /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(4247..5062)
                     /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(4247..5062)
                     /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     4970..4989
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5054..5073
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5287..5875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5776..5795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7264..7284
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggttctcgtt tcaacaagct cgacctcaat cttctggtcg ccctggatgc actgctcacg
       61 gagatgagca tcagccgcgc cgccgaaaag atccatctga gccagtcggc catgagcaat
      121 gccctggcgc ggctgcgcga gtatttcgat gatgaattgc tgatccaggt gggccggcgc
      181 atggagccca cgccgcgcgc cgaggtgctc aaggatgcgg tgcatgatgt gctgcggcgt
      241 atcgatggct ccatcgcggc gctgccggcc ttcgtgccgg ccgagtccac gcgcgagttt
      301 cgcatctcgg tttcggactt tacgctctcc gtcctcatcc cccgggtgct ggcgcgcgcg
      361 cacgccgagg gcaagcacat ccgctttgcc ctgatgccgc aggtgcaaga cccgacccgc
      421 tcgctggatc gggccgaggt ggacctgctg gtcttgccgc aggaattctg cacgcccgat
      481 catcctgccg aagaggtctt ccgcgaacgg catgtctgcg tggtctggcg cgacagtgcg
      541 ctggcgcaag gcgagctgac gctggaacgc tacatggcct caggccatgt ggtgatggtg
      601 ccgcctgggg ccaatgcgtc gtcggtggag gcgtggatgg ccaggaagct gggctttgcg
      661 cgccgggtgg aagtgaccag cttcagcttc gcttctgcgc tggcgctggt acaggggacg
      721 gaccgcatcg ccacggtgca tgcccggctg gcgcagctgc tggctccgca atggccggtg
      781 gtgatcaagg agagtccgct gtcgctgggc gagatgcggc agatgatgca gtggcatcgc
      841 taccgcagca atgatcctgg catccagtgg ctgcgtcggg tgtttctgga gagtgcgcag
      901 gagatggatg cggcgctgcc aggcatctgc tgaatcctaa ctcgagattg aattgaattg
      961 aaatcgatag atcaattttt ttcttttctc tttccccatc ctttacgcta aaataatagt
     1021 ttattttatt ttttgaatat tttttattta tatacgtata tatagactat tatttatctt
     1081 ttaatgatta ttaagatttt tattaaaaaa aaattcgctc ctcttttaat gcctttatgc
     1141 agtttttttt tcccattcga tatttctatg tgctgccaaa ccagatgtca acacagctac
     1201 aacgaagctt cgtcttttct tgaagaaaag aggaagctcg ctaaatggga ttccactttc
     1261 cgttccctgc cagctgatgg aaaaaggtta gtggaacgat gaagaataaa aagagagatc
     1321 cactgaggtg aaatttcagc tgacagcgag tttcatgatc gtgatgaaca atggtaacga
     1381 gttgtggctg ttgccaggga gggtggttct caccaaatcg ctacttgggt ttgagattgg
     1441 taaaattgat tgtttggatg gcaatcatct atggaatgga tattatataa caaagaagaa
     1501 ataatgaact gattctcttc ctccttcttg tcctttctta attctgttgt aattaccttc
     1561 ctttgtaatt ttttttgtaa ttattcttct taataatcca aacaaacaca catattacaa
     1621 taagatctat ggtttcaaag ggcgaagaag acaatatggc tattattaag gaattcatga
     1681 gattcaaggt ccacatggaa ggttctgtca acggtcacga attcgaaatt gaaggtgaag
     1741 gtgaaggtag accatacgaa ggtacccaaa ccgctaagtt gaaggtcacc aagggtggtc
     1801 cattgccatt cgcttgggat attttgtctc cacaattcat gtacggttct aaggcttacg
     1861 tcaagcaccc agctgatatt ccagattact tgaagttgtc tttcccagaa ggtttcaagt
     1921 gggaaagagt catgaacttc gaagatggtg gtgtcgtcac cgtcacccaa gattcttctt
     1981 tgcaagatgg tgaattcatt tacaaggtca agttgagagg taccaacttc ccatctgatg
     2041 gtccagtcat gcaaaagaag accatgggtt gggaagcttc ttctgaaaga atgtacccag
     2101 aagatggtgc tttgaagggt gaaattaagc aaagattgaa gttgaaggat ggtggtcact
     2161 acgatgctga agtcaagacc acctacaagg ctaagaagcc agtccaattg ccaggtgctt
     2221 acaacgtcaa cattaagttg gatattacct ctcacaacga agattacacc attgtcgaac
     2281 aatacgaaag agctgaaggt agacactcta ccggtggtat ggatgaattg tacaagtaaa
     2341 tcctaactcg agataaagca atcttgatga ggataatgat ttttttttga atatacataa
     2401 atactaccgt ttttctgcta gattttgtga tgacgtaaat aagtacatat tactttttaa
     2461 gccaagacaa gattaagcat taactttacc cttttctttc taagtttcaa tattagttat
     2521 cactgtttaa aagttatggc gagaacgtcg gcggttaaaa tatattaccc tgaacggctg
     2581 agcagttaca gagatgttac gaaccactag tgcactgcag tacacggttt ccttgaaatt
     2641 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     2701 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     2761 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     2821 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     2881 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     2941 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     3001 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     3061 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     3121 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     3181 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     3241 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     3301 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     3361 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     3421 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     3481 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     3541 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     3601 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     3661 tttaattata tcagttatta cccgagtaga gcacttgaat ccactgcccc gggaatctcg
     3721 gtcgtaatga tttctataat gacgaaaaaa aaaaaattgg aaagaaaaag cttcatggcc
     3781 tttataaaaa ggaactatcc aatacctcgc cagaaccaag taacagtatt ttacggggca
     3841 caaatcaaga acaataagac aggactgtaa agatggacgc attgaactcc aaagaacaac
     3901 aagagttcca aaaagtagtg gaacaaaagc aaatgaagga tttcatgcgt ttgtactcta
     3961 atctggtaga aagatgtttc acagactgtg tcaatgactt cacaacatca aagctaacca
     4021 ataaggaaca aacatgcatc atgaagtgct cagaaaagtt cttgaagcat agcgaacgtg
     4081 tagggcagcg tttccaagaa caaaacgctg ccttgggaca aggcttgggc cgataaggtg
     4141 tactggcgta tatatatcta attatgtatc tctggtgtag cccattttta gcatgtaaat
     4201 ataaagaccg agcggccgcg tgttacaacc aattaaccaa ttctgattag aaaaactcat
     4261 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     4321 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     4381 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     4441 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     4501 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     4561 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc
     4621 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     4681 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     4741 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     4801 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     4861 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     4921 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     4981 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg
     5041 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     5101 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     5161 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag tcatgaccaa
     5221 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5281 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5341 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5401 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5461 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5521 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5581 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5641 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5701 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5761 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5821 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5881 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5941 tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gccaatgcta aattcgagtg
     6001 aaacacagga agatcagaaa atcctcattt catccatatt aacaataatt tcaaatgttt
     6061 atttgcatta tttgaaacta ggcaagacaa gcaacgaaac gtttttgaaa attttgagta
     6121 ttttcaataa atttgtagag gactcagata ttgaaaaaaa gctacagcaa ttaatacttg
     6181 ataagaagag tattgagaag ggcaacggtt catcatctca tggatctgca catgaacaaa
     6241 caccagagtc aaacgacgtt gaaattgagg ctactgcgcc aattgatgac aatacagacg
     6301 atgataacaa accgaagtta tctgatgtag aaaaggatta aagatgctaa gagatagtga
     6361 tgatatttca taaataatgt aattctatat atgttaatta ccttttttgc gaggcatatt
     6421 tatggtgaag gataagtttt gaccatcaaa gaaggttaat gtggctgtgg tttcagggtc
     6481 cataaagccc acatggataa cattacccct gaattcgcat ctagactgaa ctggccgata
     6541 attgcagacg aacggtgttg ttatccgata caaccggata tttttctttt aatgagtcta
     6601 aaccgtgata gcttcaggtt aatacaatca aaaaaagctc aaatattctt ttaatgccgc
     6661 gttcacagat tccaattgaa tacaactagg tagttcatta tatgaagcct ttgctactat
     6721 ttttcactat agtctgcctt caccttaatg cagacatcca catattttaa tcactttaaa
     6781 ataaaaagga agatatatta gaagctatga tccaatctgt aagccagatt aaaattcacg
     6841 aactcttctt tcatttgaat tgaatgcttt gagttggggt agattatcgc aaattactca
     6901 tcacatttat tgactacgaa cttgctgatg tccttttttt atttatattt ttcttcagtg
     6961 aagcgatttt ttttttacac agaccaagac ggaaaaaagt agctaaggaa gaaaacaaaa
     7021 tcatgaaaaa aatgtgaagt gatcatgcac atcgcatcaa cttaaacatt ggcttagaga
     7081 tatatagagt tagagtttac ggcaaccttt aagcaccaat accttttggc atagtctaaa
     7141 gacctggttc ttaattttaa acaaatttaa ctaaagattt ccctatcaaa gaagtaacga
     7201 gttgacagat tttctcaaaa taaatcgata ctgcatttct aggcatatcc agcgagatct
     7261 atgccaaaga agaagagaaa ggtt
//
