LOCUS       sequence-266896-        7221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV2-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 7221)
  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 7221)
  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_MV2
FEATURES             Location/Qualifiers
     source          1..7221
                     /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"
     rep_origin      3718..4700
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4758..5573)
                     /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(4758..5573)
                     /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     5481..5500
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5565..5584
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5798..6386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6287..6306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6555..7221
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     7091..7114
                     /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 cccgagtaac
     3721 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     3781 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     3841 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     3901 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     3961 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     4021 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     4081 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     4141 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     4201 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     4261 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     4321 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     4381 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     4441 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     4501 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     4561 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     4621 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     4681 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaac
     4741 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     4801 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     4861 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     4921 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     4981 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag
     5041 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     5101 ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa
     5161 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     5221 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     5281 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     5341 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     5401 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     5461 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     5521 ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct cataacaccc
     5581 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     5641 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     5701 tttgctgagt tgaaggatca gtcatgacca aaatccctta acgtgagttt tcgttccact
     5761 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5821 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5881 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5941 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6001 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6061 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6121 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6181 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6241 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6301 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6361 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6421 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6481 accgtgcggc cgcccctgaa ttcgcatcta gactgaactg gccgataatt gcagacgaac
     6541 gcagttcgag tttatcatta tcaatactgc catttcaaag aatacgtaaa taattaatag
     6601 tagtgatttt cctaacttta tttagtcaaa aaattagcct tttaattctg ctgtaacccg
     6661 tacatgccca aaataggggg cgggttacac agaatatata acatcgtagg tgtctgggtg
     6721 aacagtttat tcctggcatc cactaaatat aatggagccc gctttttaag ctggcatcca
     6781 gaaaaaaaaa gaatcccagc accaaaatat tgttttcttc accaaccatc agttcatagg
     6841 tccattctct tagcgcaact acagagaaca ggggcacaaa caggcaaaaa acgggcacaa
     6901 cctcaatgga gtgatgcaac ctgcctggag taaatgatga cacaaggcaa ttgacccacg
     6961 catgtatcta tctcattttc ttacaccttc tattaccttc tgctctctct gatttggaaa
     7021 aagctgaaaa aaaaggttga aaccagttcc ctgaaattat tcccctactt gactaataag
     7081 tatataaaga cggtaggtat tgattgtaat tctgtaaatc tatttcttaa acttcttaaa
     7141 ttctactttt atagttagtc ttttttttag ttttaaaaca ccaagaactt agtttcgaat
     7201 aaacacacat aaacaaacaa a
//
