LOCUS       sequence-266111-        6810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-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 6810)
  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 6810)
  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_MV1
FEATURES             Location/Qualifiers
     source          1..6810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..667
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     537..560
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             677..697
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1668..1688
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1692..1712
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3581..4384
                     /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    4467..4970
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(5014..5829)
                     /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(5014..5829)
                     /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     5737..5756
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5821..5840
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6054..6642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6543..6562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcattatcaa tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta
       61 actttattta gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat
      121 agggggcggg ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct
      181 ggcatccact aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat
      241 cccagcacca aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc
      301 gcaactacag agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga
      361 tgcaacctgc ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc
      421 attttcttac accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa
      481 ggttgaaacc agttccctga aattattccc ctacttgact aataagtata taaagacggt
      541 aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag
      601 ttagtctttt ttttagtttt aaaacaccaa gaacttagtt tcgaataaac acacataaac
      661 aaacaaaaga tctatgccaa agaagaagag aaaggttggt tctcgtttca acaagctcga
      721 cctcaatctt ctggtcgccc tggatgcact gctcacggag atgagcatca gccgcgccgc
      781 cgaaaagatc catctgagcc agtcggccat gagcaatgcc ctggcgcggc tgcgcgagta
      841 tttcgatgat gaattgctga tccaggtggg ccggcgcatg gagcccacgc cgcgcgccga
      901 ggtgctcaag gatgcggtgc atgatgtgct gcggcgtatc gatggctcca tcgcggcgct
      961 gccggccttc gtgccggccg agtccacgcg cgagtttcgc atctcggttt cggactttac
     1021 gctctccgtc ctcatccccc gggtgctggc gcgcgcgcac gccgagggca agcacatccg
     1081 ctttgccctg atgccgcagg tgcaagaccc gacccgctcg ctggatcggg ccgaggtgga
     1141 cctgctggtc ttgccgcagg aattctgcac gcccgatcat cctgccgaag aggtcttccg
     1201 cgaacggcat gtctgcgtgg tctggcgcga cagtgcgctg gcgcaaggcg agctgacgct
     1261 ggaacgctac atggcctcag gccatgtggt gatggtgccg cctggggcca atgcgtcgtc
     1321 ggtggaggcg tggatggcca ggaagctggg ctttgcgcgc cgggtggaag tgaccagctt
     1381 cagcttcgct tctgcgctgg cgctggtaca ggggacggac cgcatcgcca cggtgcatgc
     1441 ccggctggcg cagctgctgg ctccgcaatg gccggtggtg atcaaggaga gtccgctgtc
     1501 gctgggcgag atgcggcaga tgatgcagtg gcatcgctac cgcagcaatg atcctggcat
     1561 ccagtggctg cgtcgggtgt ttctggagag tgcgcaggag atggatgcgg cgctgccagg
     1621 catctgcgga tcctgagacc aggctgccag gcatctgcgg atccgatcca aaaaagaaga
     1681 gaaaggtaga tccaaaaaag aagagaaagg tataactcga gtggcattga attgaattga
     1741 aatcgataga tcaatttttt tcttttctct ttccccatcc tttacgctaa aataatagtt
     1801 tattttattt tttgaatatt ttttatttat atacgtatat atagactatt atttatcttt
     1861 taatgattat taagattttt attaaaaaaa aattcgctcc tcttttaatg cctttatgca
     1921 gttttttttt cccattcgat atttctatgt gctgccaaac cagatgtcaa cacagctaca
     1981 acgaagcttc gtcttttctt gaagaaaaga ggaagctcgc taaatgggat tccactttcc
     2041 gttccctgcc agctgatgga aaaaggttag tggaacgatg aagaataaaa agagagatcc
     2101 actgaggtga aatttcagct gacagcgagt ttcatgatcg tgatgaacaa tggtaacgag
     2161 ttgtggctgt tgccagggag ggtggttctc accaaatcgc tacttgggtt tgagattggt
     2221 aaaattgatt gtttggatgg caatcatcta tggaatggat attatataac aaagaagaaa
     2281 taatgaactg attctcttcc tccttcttgt cctttcttaa ttctgttgta attaccttcc
     2341 tttgtaattt tttttgtaat tattcttctt aataatccaa acaaacacac atattacaat
     2401 aagatctatg gtttcaaagg gcgaagaaga caatatggct attattaagg aattcatgag
     2461 attcaaggtc cacatggaag gttctgtcaa cggtcacgaa ttcgaaattg aaggtgaagg
     2521 tgaaggtaga ccatacgaag gtacccaaac cgctaagttg aaggtcacca agggtggtcc
     2581 attgccattc gcttgggata ttttgtctcc acaattcatg tacggttcta aggcttacgt
     2641 caagcaccca gctgatattc cagattactt gaagttgtct ttcccagaag gtttcaagtg
     2701 ggaaagagtc atgaacttcg aagatggtgg tgtcgtcacc gtcacccaag attcttcttt
     2761 gcaagatggt gaattcattt acaaggtcaa gttgagaggt accaacttcc catctgatgg
     2821 tccagtcatg caaaagaaga ccatgggttg ggaagcttct tctgaaagaa tgtacccaga
     2881 agatggtgct ttgaagggtg aaattaagca aagattgaag ttgaaggatg gtggtcacta
     2941 cgatgctgaa gtcaagacca cctacaaggc taagaagcca gtccaattgc caggtgctta
     3001 caacgtcaac attaagttgg atattacctc tcacaacgaa gattacacca ttgtcgaaca
     3061 atacgaaaga gctgaaggta gacactctac cggtggtatg gatgaattgt acaagtaaat
     3121 cctaactcga gataaagcaa tcttgatgag gataatgatt tttttttgaa tatacataaa
     3181 tactaccgtt tttctgctag attttgtgat gacgtaaata agtacatatt actttttaag
     3241 ccaagacaag attaagcatt aactttaccc ttttctttct aagtttcaat attagttatc
     3301 actgtttaaa agttatggcg agaacgtcgg cggttaaaat atattaccct gaacggctga
     3361 gcagttacag agatgttacg aaccactagt gcactgcagt acacggtttc cttgaaattt
     3421 ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     3481 ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     3541 actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     3601 ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     3661 aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     3721 agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     3781 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     3841 actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     3901 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     3961 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     4021 tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     4081 tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     4141 catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     4201 tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg
     4261 atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     4321 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     4381 ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     4441 ttaattatat cagttattac ccgagtatca cgtgctataa aaataattat aatttaaatt
     4501 ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata
     4561 gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt
     4621 atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa
     4681 aaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct
     4741 atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt
     4801 ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta
     4861 ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt
     4921 attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtcccgagc
     4981 ggccgcgtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     5041 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     5101 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     5161 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     5221 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     5281 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     5341 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc
     5401 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     5461 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     5521 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     5581 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     5641 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     5701 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     5761 taaatcagca tccatgttgg aatttaatcg cggcctggag caagacgttt cccgttgaat
     5821 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     5881 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg
     5941 ttgaataaat cgaacttttg ctgagttgaa ggatcagtca tgaccaaaat cccttaacgt
     6001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6721 cctgattctg tggataaccg tgcggccgcc cctgaattcg catctagact gaactggccg
     6781 ataattgcag acgaacgcag ttcgagttta
//
