LOCUS       sequence-266873-        7303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV2-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 7303)
  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 7303)
  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_MV2
FEATURES             Location/Qualifiers
     source          1..7303
                     /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             1001..1021
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1025..1045
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2914..3717
                     /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"
     rep_origin      3800..4782
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4840..5655)
                     /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(4840..5655)
                     /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     5563..5582
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5647..5666
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5880..6468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6369..6388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6637..7303
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     7173..7196
                     /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 ggatcctgag accaggctgc caggcatctg cggatccgat ccaaaaaaga agagaaaggt
     1021 agatccaaaa aagaagagaa aggtataact cgagtggcat tgaattgaat tgaaatcgat
     1081 agatcaattt ttttcttttc tctttcccca tcctttacgc taaaataata gtttatttta
     1141 ttttttgaat attttttatt tatatacgta tatatagact attatttatc ttttaatgat
     1201 tattaagatt tttattaaaa aaaaattcgc tcctctttta atgcctttat gcagtttttt
     1261 tttcccattc gatatttcta tgtgctgcca aaccagatgt caacacagct acaacgaagc
     1321 ttcgtctttt cttgaagaaa agaggaagct cgctaaatgg gattccactt tccgttccct
     1381 gccagctgat ggaaaaaggt tagtggaacg atgaagaata aaaagagaga tccactgagg
     1441 tgaaatttca gctgacagcg agtttcatga tcgtgatgaa caatggtaac gagttgtggc
     1501 tgttgccagg gagggtggtt ctcaccaaat cgctacttgg gtttgagatt ggtaaaattg
     1561 attgtttgga tggcaatcat ctatggaatg gatattatat aacaaagaag aaataatgaa
     1621 ctgattctct tcctccttct tgtcctttct taattctgtt gtaattacct tcctttgtaa
     1681 ttttttttgt aattattctt cttaataatc caaacaaaca cacatattac aataagatct
     1741 atggtttcaa agggcgaaga agacaatatg gctattatta aggaattcat gagattcaag
     1801 gtccacatgg aaggttctgt caacggtcac gaattcgaaa ttgaaggtga aggtgaaggt
     1861 agaccatacg aaggtaccca aaccgctaag ttgaaggtca ccaagggtgg tccattgcca
     1921 ttcgcttggg atattttgtc tccacaattc atgtacggtt ctaaggctta cgtcaagcac
     1981 ccagctgata ttccagatta cttgaagttg tctttcccag aaggtttcaa gtgggaaaga
     2041 gtcatgaact tcgaagatgg tggtgtcgtc accgtcaccc aagattcttc tttgcaagat
     2101 ggtgaattca tttacaaggt caagttgaga ggtaccaact tcccatctga tggtccagtc
     2161 atgcaaaaga agaccatggg ttgggaagct tcttctgaaa gaatgtaccc agaagatggt
     2221 gctttgaagg gtgaaattaa gcaaagattg aagttgaagg atggtggtca ctacgatgct
     2281 gaagtcaaga ccacctacaa ggctaagaag ccagtccaat tgccaggtgc ttacaacgtc
     2341 aacattaagt tggatattac ctctcacaac gaagattaca ccattgtcga acaatacgaa
     2401 agagctgaag gtagacactc taccggtggt atggatgaat tgtacaagta aatcctaact
     2461 cgagataaag caatcttgat gaggataatg attttttttt gaatatacat aaatactacc
     2521 gtttttctgc tagattttgt gatgacgtaa ataagtacat attacttttt aagccaagac
     2581 aagattaagc attaacttta cccttttctt tctaagtttc aatattagtt atcactgttt
     2641 aaaagttatg gcgagaacgt cggcggttaa aatatattac cctgaacggc tgagcagtta
     2701 cagagatgtt acgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat
     2761 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     2821 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     2881 agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     2941 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     3001 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     3061 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     3121 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     3181 gaggacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     3241 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     3301 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     3361 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     3421 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     3481 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     3541 ggagatgcat tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac
     3601 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     3661 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     3721 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     3781 tatcagttat tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     3841 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     3901 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     3961 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     4021 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     4081 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     4141 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     4201 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     4261 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     4321 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     4381 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     4441 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     4501 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     4561 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     4621 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     4681 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
     4741 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
     4801 ccgagcggcc gcgtgttaca accaattaac caattctgat tagaaaaact catcgagcat
     4861 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     4921 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     4981 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     5041 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     5101 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     5161 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac
     5221 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     5281 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     5341 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     5401 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     5461 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     5521 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     5581 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg
     5641 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     5701 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     5761 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagtcatgac caaaatccct
     5821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6001 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6541 ttatcccctg attctgtgga taaccgtgcg gccgcccctg aattcgcatc tagactgaac
     6601 tggccgataa ttgcagacga acgcagttcg agtttatcat tatcaatact gccatttcaa
     6661 agaatacgta aataattaat agtagtgatt ttcctaactt tatttagtca aaaaattagc
     6721 cttttaattc tgctgtaacc cgtacatgcc caaaataggg ggcgggttac acagaatata
     6781 taacatcgta ggtgtctggg tgaacagttt attcctggca tccactaaat ataatggagc
     6841 ccgcttttta agctggcatc cagaaaaaaa aagaatccca gcaccaaaat attgttttct
     6901 tcaccaacca tcagttcata ggtccattct cttagcgcaa ctacagagaa caggggcaca
     6961 aacaggcaaa aaacgggcac aacctcaatg gagtgatgca acctgcctgg agtaaatgat
     7021 gacacaaggc aattgaccca cgcatgtatc tatctcattt tcttacacct tctattacct
     7081 tctgctctct ctgatttgga aaaagctgaa aaaaaaggtt gaaaccagtt ccctgaaatt
     7141 attcccctac ttgactaata agtatataaa gacggtaggt attgattgta attctgtaaa
     7201 tctatttctt aaacttctta aattctactt ttatagttag tctttttttt agttttaaaa
     7261 caccaagaac ttagtttcga ataaacacac ataaacaaac aaa
//
