LOCUS       sequence-266880-        6728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pTDH3_NLS_FdeR-N_tPGK1-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6728)
  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 6728)
  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_001_026_MV1
FEATURES             Location/Qualifiers
     source          1..6728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..30
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2832..3635
                     /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    3718..4221
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4265..5080)
                     /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(4265..5080)
                     /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     4988..5007
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5072..5091
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5305..5893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5794..5813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6062..6728
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     6598..6621
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
ORIGIN
        1 agatctatgc caaagaagaa gagaaaggtt ggttctcgtt tcaacaagct cgacctcaat
       61 cttctggtcg ccctggatgc actgctcacg gagatgagca tcagccgcgc cgccgaaaag
      121 atccatctga gccagtcggc catgagcaat gccctggcgc ggctgcgcga gtatttcgat
      181 gatgaattgc tgatccaggt gggccggcgc atggagccca cgccgcgcgc cgaggtgctc
      241 aaggatgcgg tgcatgatgt gctgcggcgt atcgatggct ccatcgcggc gctgccggcc
      301 ttcgtgccgg ccgagtccac gcgcgagttt cgcatctcgg tttcggactt tacgctctcc
      361 gtcctcatcc cccgggtgct ggcgcgcgcg cacgccgagg gcaagcacat ccgctttgcc
      421 ctgatgccgc aggtgcaaga cccgacccgc tcgctggatc gggccgaggt ggacctgctg
      481 gtcttgccgc aggaattctg cacgcccgat catcctgccg aagaggtctt ccgcgaacgg
      541 catgtctgcg tggtctggcg cgacagtgcg ctggcgcaag gcgagctgac gctggaacgc
      601 tacatggcct caggccatgt ggtgatggtg ccgcctgggg ccaatgcgtc gtcggtggag
      661 gcgtggatgg ccaggaagct gggctttgcg cgccgggtgg aagtgaccag cttcagcttc
      721 gcttctgcgc tggcgctggt acaggggacg gaccgcatcg ccacggtgca tgcccggctg
      781 gcgcagctgc tggctccgca atggccggtg gtgatcaagg agagtccgct gtcgctgggc
      841 gagatgcggc agatgatgca gtggcatcgc taccgcagca atgatcctgg catccagtgg
      901 ctgcgtcggg tgtttctgga gagtgcgcag gagatggatg cggcgctgcc aggcatctgc
      961 tgaatcctaa ctcgagattg aattgaattg aaatcgatag atcaattttt ttcttttctc
     1021 tttccccatc ctttacgcta aaataatagt ttattttatt ttttgaatat tttttattta
     1081 tatacgtata tatagactat tatttatctt ttaatgatta ttaagatttt tattaaaaaa
     1141 aaattcgctc ctcttttaat gcctttatgc agtttttttt tcccattcga tatttctatg
     1201 tgctgccaaa ccagatgtca acacagctac aacgaagctt cgtcttttct tgaagaaaag
     1261 aggaagctcg ctaaatggga ttccactttc cgttccctgc cagctgatgg aaaaaggtta
     1321 gtggaacgat gaagaataaa aagagagatc cactgaggtg aaatttcagc tgacagcgag
     1381 tttcatgatc gtgatgaaca atggtaacga gttgtggctg ttgccaggga gggtggttct
     1441 caccaaatcg ctacttgggt ttgagattgg taaaattgat tgtttggatg gcaatcatct
     1501 atggaatgga tattatataa caaagaagaa ataatgaact gattctcttc ctccttcttg
     1561 tcctttctta attctgttgt aattaccttc ctttgtaatt ttttttgtaa ttattcttct
     1621 taataatcca aacaaacaca catattacaa taagatctat ggtttcaaag ggcgaagaag
     1681 acaatatggc tattattaag gaattcatga gattcaaggt ccacatggaa ggttctgtca
     1741 acggtcacga attcgaaatt gaaggtgaag gtgaaggtag accatacgaa ggtacccaaa
     1801 ccgctaagtt gaaggtcacc aagggtggtc cattgccatt cgcttgggat attttgtctc
     1861 cacaattcat gtacggttct aaggcttacg tcaagcaccc agctgatatt ccagattact
     1921 tgaagttgtc tttcccagaa ggtttcaagt gggaaagagt catgaacttc gaagatggtg
     1981 gtgtcgtcac cgtcacccaa gattcttctt tgcaagatgg tgaattcatt tacaaggtca
     2041 agttgagagg taccaacttc ccatctgatg gtccagtcat gcaaaagaag accatgggtt
     2101 gggaagcttc ttctgaaaga atgtacccag aagatggtgc tttgaagggt gaaattaagc
     2161 aaagattgaa gttgaaggat ggtggtcact acgatgctga agtcaagacc acctacaagg
     2221 ctaagaagcc agtccaattg ccaggtgctt acaacgtcaa cattaagttg gatattacct
     2281 ctcacaacga agattacacc attgtcgaac aatacgaaag agctgaaggt agacactcta
     2341 ccggtggtat ggatgaattg tacaagtaaa tcctaactcg agataaagca atcttgatga
     2401 ggataatgat ttttttttga atatacataa atactaccgt ttttctgcta gattttgtga
     2461 tgacgtaaat aagtacatat tactttttaa gccaagacaa gattaagcat taactttacc
     2521 cttttctttc taagtttcaa tattagttat cactgtttaa aagttatggc gagaacgtcg
     2581 gcggttaaaa tatattaccc tgaacggctg agcagttaca gagatgttac gaaccactag
     2641 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     2701 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     2761 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     2821 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     2881 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     2941 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     3001 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     3061 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     3121 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     3181 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     3241 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     3301 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     3361 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     3421 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     3481 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     3541 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     3601 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     3661 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtatc
     3721 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     3781 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     3841 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     3901 tgccgaattg tttcatcttg tctgtgtaga aaaccacaca cgaaaatcct gtgattttac
     3961 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     4021 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     4081 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     4141 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     4201 gtaagttaca ggcaagcgat ccgtcccgag cggccgcgtg ttacaaccaa ttaaccaatt
     4261 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     4321 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4381 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4441 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4501 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     4561 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4621 attgcgcctg agcgaggcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4681 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4741 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     4801 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4861 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4921 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     4981 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     5041 gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     5101 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     5161 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     5221 aggatcagtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5521 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5581 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5641 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5701 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5761 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5821 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5881 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5941 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtgcggccgc
     6001 ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgca gttcgagttt
     6061 atcattatca atactgccat ttcaaagaat acgtaaataa ttaatagtag tgattttcct
     6121 aactttattt agtcaaaaaa ttagcctttt aattctgctg taacccgtac atgcccaaaa
     6181 tagggggcgg gttacacaga atatataaca tcgtaggtgt ctgggtgaac agtttattcc
     6241 tggcatccac taaatataat ggagcccgct ttttaagctg gcatccagaa aaaaaaagaa
     6301 tcccagcacc aaaatattgt tttcttcacc aaccatcagt tcataggtcc attctcttag
     6361 cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct caatggagtg
     6421 atgcaacctg cctggagtaa atgatgacac aaggcaattg acccacgcat gtatctatct
     6481 cattttctta caccttctat taccttctgc tctctctgat ttggaaaaag ctgaaaaaaa
     6541 aggttgaaac cagttccctg aaattattcc cctacttgac taataagtat ataaagacgg
     6601 taggtattga ttgtaattct gtaaatctat ttcttaaact tcttaaattc tacttttata
     6661 gttagtcttt tttttagttt taaaacacca agaacttagt ttcgaataaa cacacataaa
     6721 caaacaaa
//
