LOCUS       sequence-258377-        6779 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 6779)
  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 6779)
  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..6779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(40..855)
                     /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(40..855)
                     /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     763..782
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     847..866
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1080..1668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1569..1588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1837..2503
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2373..2396
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2513..2533
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3473..3493
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3497..3517
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5386..6189
                     /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    6272..6775
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ccgagcggcc gcgtgttaca accaattaac caattctgat tagaaaaact catcgagcat
       61 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
      121 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
      181 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
      241 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
      301 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
      361 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac
      421 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
      481 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
      541 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
      601 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
      661 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
      721 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
      781 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg
      841 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
      901 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
      961 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagtcatgac caaaatccct
     1021 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1741 ttatcccctg attctgtgga taaccgtgcg gccgcccctg aattcgcatc tagactgaac
     1801 tggccgataa ttgcagacga acgcagttcg agtttatcat tatcaatact gccatttcaa
     1861 agaatacgta aataattaat agtagtgatt ttcctaactt tatttagtca aaaaattagc
     1921 cttttaattc tgctgtaacc cgtacatgcc caaaataggg ggcgggttac acagaatata
     1981 taacatcgta ggtgtctggg tgaacagttt attcctggca tccactaaat ataatggagc
     2041 ccgcttttta agctggcatc cagaaaaaaa aagaatccca gcaccaaaat attgttttct
     2101 tcaccaacca tcagttcata ggtccattct cttagcgcaa ctacagagaa caggggcaca
     2161 aacaggcaaa aaacgggcac aacctcaatg gagtgatgca acctgcctgg agtaaatgat
     2221 gacacaaggc aattgaccca cgcatgtatc tatctcattt tcttacacct tctattacct
     2281 tctgctctct ctgatttgga aaaagctgaa aaaaaaggtt gaaaccagtt ccctgaaatt
     2341 attcccctac ttgactaata agtatataaa gacggtaggt attgattgta attctgtaaa
     2401 tctatttctt aaacttctta aattctactt ttatagttag tctttttttt agttttaaaa
     2461 caccaagaac ttagtttcga ataaacacac ataaacaaac aaaagatcta tgccaaagaa
     2521 gaagagaaag gttggttctc gtttcaacaa gctcgacctc aatcttctgg tcgccctgga
     2581 tgcactgctc acggagatga gcatcagccg cgccgccgaa aagatccatc tgagccagtc
     2641 ggccatgagc aatgccctgg cgcggctgcg cgagtatttc gatgatgaat tgctgatcca
     2701 ggtgggccgg cgcatggagc ccacgccgcg cgccgaggtg ctcaaggatg cggtgcatga
     2761 tgtgctgcgg cgtatcgatg gctccatcgc ggcgctgccg gccttcgtgc cggccgagtc
     2821 cacgcgcgag tttcgcatct cggtttcgga ctttacgctc tccgtcctca tcccccgggt
     2881 gctggcgcgc gcgcacgccg agggcaagca catccgcttt gccctgatgc cgcaggtgca
     2941 agacccgacc cgctcgctgg atcgggccga ggtggacctg ctggtcttgc cgcaggaatt
     3001 ctgcacgccc gatcatcctg ccgaagaggt cttccgcgaa cggcatgtct gcgtggtctg
     3061 gcgcgacagt gcgctggcgc aaggcgagct gacgctggaa cgctacatgg cctcaggcca
     3121 tgtggtgatg gtgccgcctg gggccaatgc gtcgtcggtg gaggcgtgga tggccaggaa
     3181 gctgggcttt gcgcgccggg tggaagtgac cagcttcagc ttcgcttctg cgctggcgct
     3241 ggtacagggg acggaccgca tcgccacggt gcatgcccgg ctggcgcagc tgctggctcc
     3301 gcaatggccg gtggtgatca aggagagtcc gctgtcgctg ggcgagatgc ggcagatgat
     3361 gcagtggcat cgctaccgca gcaatgatcc tggcatccag tggctgcgtc gggtgtttct
     3421 ggagagtgcg caggagatgg atgcggcgct gccaggcatc tgcggatccg atccaaaaaa
     3481 gaagagaaag gtagatccaa aaaagaagag aaaggtataa ctcgagtggc attgaattga
     3541 attgaaatcg atagatcaat ttttttcttt tctctttccc catcctttac gctaaaataa
     3601 tagtttattt tattttttga atatttttta tttatatacg tatatataga ctattattta
     3661 tcttttaatg attattaaga tttttattaa aaaaaaattc gctcctcttt taatgccttt
     3721 atgcagtttt tttttcccat tcgatatttc tatgtgctgc caaaccagat gtcaacacag
     3781 ctacaacgaa gcttcgtctt ttcttgaaga aaagaggaag ctcgctaaat gggattccac
     3841 tttccgttcc ctgccagctg atggaaaaag gttagtggaa cgatgaagaa taaaaagaga
     3901 gatccactga ggtgaaattt cagctgacag cgagtttcat gatcgtgatg aacaatggta
     3961 acgagttgtg gctgttgcca gggagggtgg ttctcaccaa atcgctactt gggtttgaga
     4021 ttggtaaaat tgattgtttg gatggcaatc atctatggaa tggatattat ataacaaaga
     4081 agaaataatg aactgattct cttcctcctt cttgtccttt cttaattctg ttgtaattac
     4141 cttcctttgt aatttttttt gtaattattc ttcttaataa tccaaacaaa cacacatatt
     4201 acaataagat ctatggtttc aaagggcgaa gaagacaata tggctattat taaggaattc
     4261 atgagattca aggtccacat ggaaggttct gtcaacggtc acgaattcga aattgaaggt
     4321 gaaggtgaag gtagaccata cgaaggtacc caaaccgcta agttgaaggt caccaagggt
     4381 ggtccattgc cattcgcttg ggatattttg tctccacaat tcatgtacgg ttctaaggct
     4441 tacgtcaagc acccagctga tattccagat tacttgaagt tgtctttccc agaaggtttc
     4501 aagtgggaaa gagtcatgaa cttcgaagat ggtggtgtcg tcaccgtcac ccaagattct
     4561 tctttgcaag atggtgaatt catttacaag gtcaagttga gaggtaccaa cttcccatct
     4621 gatggtccag tcatgcaaaa gaagaccatg ggttgggaag cttcttctga aagaatgtac
     4681 ccagaagatg gtgctttgaa gggtgaaatt aagcaaagat tgaagttgaa ggatggtggt
     4741 cactacgatg ctgaagtcaa gaccacctac aaggctaaga agccagtcca attgccaggt
     4801 gcttacaacg tcaacattaa gttggatatt acctctcaca acgaagatta caccattgtc
     4861 gaacaatacg aaagagctga aggtagacac tctaccggtg gtatggatga attgtacaag
     4921 taaatcctaa ctcgagataa agcaatcttg atgaggataa tgattttttt ttgaatatac
     4981 ataaatacta ccgtttttct gctagatttt gtgatgacgt aaataagtac atattacttt
     5041 ttaagccaag acaagattaa gcattaactt tacccttttc tttctaagtt tcaatattag
     5101 ttatcactgt ttaaaagtta tggcgagaac gtcggcggtt aaaatatatt accctgaacg
     5161 gctgagcagt tacagagatg ttacgaacca ctagtgcact gcagtacacg gtttccttga
     5221 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     5281 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     5341 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     5401 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     5461 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     5521 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     5581 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     5641 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     5701 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     5761 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     5821 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     5881 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     5941 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     6001 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     6061 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     6121 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     6181 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     6241 tcaatttaat tatatcagtt attacccgag tatcacgtgc tataaaaata attataattt
     6301 aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa
     6361 aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac
     6421 cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg
     6481 tagaaaacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta
     6541 tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg
     6601 aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt
     6661 atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc
     6721 aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtc
//
