LOCUS       sequence-266884-        6772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pRPL18B_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 6772)
  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 6772)
  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_005_026_MV1
FEATURES             Location/Qualifiers
     source          1..6772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             949..969
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             973..993
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2862..3665
                     /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    3748..4251
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4295..5110)
                     /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(4295..5110)
                     /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     5018..5037
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5102..5121
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5335..5923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5824..5843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacaaaagat ctatgcgttt caacaagctc gacctcaatc ttctggtcgc cctggatgca
       61 ctgctcacgg agatgagcat cagccgcgcc gccgaaaaga tccatctgag ccagtcggcc
      121 atgagcaatg ccctggcgcg gctgcgcgag tatttcgatg atgaattgct gatccaggtg
      181 ggccggcgca tggagcccac gccgcgcgcc gaggtgctca aggatgcggt gcatgatgtg
      241 ctgcggcgta tcgatggctc catcgcggcg ctgccggcct tcgtgccggc cgagtccacg
      301 cgcgagtttc gcatctcggt ttcggacttt acgctctccg tcctcatccc ccgggtgctg
      361 gcgcgcgcgc acgccgaggg caagcacatc cgctttgccc tgatgccgca ggtgcaagac
      421 ccgacccgct cgctggatcg ggccgaggtg gacctgctgg tcttgccgca ggaattctgc
      481 acgcccgatc atcctgccga agaggtcttc cgcgaacggc atgtctgcgt ggtctggcgc
      541 gacagtgcgc tggcgcaagg cgagctgacg ctggaacgct acatggcctc aggccatgtg
      601 gtgatggtgc cgcctggggc caatgcgtcg tcggtggagg cgtggatggc caggaagctg
      661 ggctttgcgc gccgggtgga agtgaccagc ttcagcttcg cttctgcgct ggcgctggta
      721 caggggacgg accgcatcgc cacggtgcat gcccggctgg cgcagctgct ggctccgcaa
      781 tggccggtgg tgatcaagga gagtccgctg tcgctgggcg agatgcggca gatgatgcag
      841 tggcatcgct accgcagcaa tgatcctggc atccagtggc tgcgtcgggt gtttctggag
      901 agtgcgcagg agatggatgc ggcgctgcca ggcatctgcg gatccgatcc aaaaaagaag
      961 agaaaggtag atccaaaaaa gaagagaaag gtataactcg agtggcattg aattgaattg
     1021 aaatcgatag atcaattttt ttcttttctc tttccccatc ctttacgcta aaataatagt
     1081 ttattttatt ttttgaatat tttttattta tatacgtata tatagactat tatttatctt
     1141 ttaatgatta ttaagatttt tattaaaaaa aaattcgctc ctcttttaat gcctttatgc
     1201 agtttttttt tcccattcga tatttctatg tgctgccaaa ccagatgtca acacagctac
     1261 aacgaagctt cgtcttttct tgaagaaaag aggaagctcg ctaaatggga ttccactttc
     1321 cgttccctgc cagctgatgg aaaaaggtta gtggaacgat gaagaataaa aagagagatc
     1381 cactgaggtg aaatttcagc tgacagcgag tttcatgatc gtgatgaaca atggtaacga
     1441 gttgtggctg ttgccaggga gggtggttct caccaaatcg ctacttgggt ttgagattgg
     1501 taaaattgat tgtttggatg gcaatcatct atggaatgga tattatataa caaagaagaa
     1561 ataatgaact gattctcttc ctccttcttg tcctttctta attctgttgt aattaccttc
     1621 ctttgtaatt ttttttgtaa ttattcttct taataatcca aacaaacaca catattacaa
     1681 taagatctat ggtttcaaag ggcgaagaag acaatatggc tattattaag gaattcatga
     1741 gattcaaggt ccacatggaa ggttctgtca acggtcacga attcgaaatt gaaggtgaag
     1801 gtgaaggtag accatacgaa ggtacccaaa ccgctaagtt gaaggtcacc aagggtggtc
     1861 cattgccatt cgcttgggat attttgtctc cacaattcat gtacggttct aaggcttacg
     1921 tcaagcaccc agctgatatt ccagattact tgaagttgtc tttcccagaa ggtttcaagt
     1981 gggaaagagt catgaacttc gaagatggtg gtgtcgtcac cgtcacccaa gattcttctt
     2041 tgcaagatgg tgaattcatt tacaaggtca agttgagagg taccaacttc ccatctgatg
     2101 gtccagtcat gcaaaagaag accatgggtt gggaagcttc ttctgaaaga atgtacccag
     2161 aagatggtgc tttgaagggt gaaattaagc aaagattgaa gttgaaggat ggtggtcact
     2221 acgatgctga agtcaagacc acctacaagg ctaagaagcc agtccaattg ccaggtgctt
     2281 acaacgtcaa cattaagttg gatattacct ctcacaacga agattacacc attgtcgaac
     2341 aatacgaaag agctgaaggt agacactcta ccggtggtat ggatgaattg tacaagtaaa
     2401 tcctaactcg agataaagca atcttgatga ggataatgat ttttttttga atatacataa
     2461 atactaccgt ttttctgcta gattttgtga tgacgtaaat aagtacatat tactttttaa
     2521 gccaagacaa gattaagcat taactttacc cttttctttc taagtttcaa tattagttat
     2581 cactgtttaa aagttatggc gagaacgtcg gcggttaaaa tatattaccc tgaacggctg
     2641 agcagttaca gagatgttac gaaccactag tgcactgcag tacacggttt ccttgaaatt
     2701 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     2761 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     2821 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     2881 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     2941 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     3001 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     3061 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     3121 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     3181 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     3241 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     3301 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     3361 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     3421 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     3481 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     3541 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     3601 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     3661 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     3721 tttaattata tcagttatta cccgagtatc acgtgctata aaaataatta taatttaaat
     3781 tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat
     3841 agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt
     3901 tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga
     3961 aaaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc
     4021 tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat
     4081 tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt
     4141 actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat
     4201 tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtcccgag
     4261 cggccgcgtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat
     4321 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     4381 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     4441 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     4501 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct
     4561 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     4621 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgaggcga aatacgcgat
     4681 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     4741 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     4801 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     4861 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     4921 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     4981 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     5041 ataaatcagc atccatgttg gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa
     5101 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     5161 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
     5221 gttgaataaa tcgaactttt gctgagttga aggatcagtc atgaccaaaa tcccttaacg
     5281 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5461 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6001 ccctgattct gtggataacc gtgcggccgc ccctgaattc gcatctagac tgaactggcc
     6061 gataattgca gacgaacgaa gaggatgtcc aatatttttt ttaaggaata aggatacttc
     6121 aagactagat tcccccctgc attcccatca gaaccgtaaa ccttggcgct ttccttggga
     6181 agtattcaag aagtgccttg tccggtttct gtggctcaca aaccagcgcg cccgatatgg
     6241 ctttcttttc acttatgaat gtaccagtac gggacaatta gaacgctcct gtaacaatct
     6301 ctttgcaaat gtggggttac attctaacca tgtcacactg ctgacgaaat tcaaagtaaa
     6361 aaaaaatggg accacgtctt gagaacgata gattttcttt attttacatt gaacagtcgt
     6421 tgtctcagcg cgctttatgt tttcattcat acttcatatt ataaaataac aaaagaagaa
     6481 tttcatattc acgcccaaga aatcaggctg ctttccaaat gcaattgaca cttcattagc
     6541 catcacacaa aactctttct tgctggagct tcttttaaaa aagacctcag tacaccaaac
     6601 acgttacccg acctcgttat tttacgacaa ctatgataaa attctgaaga aaaaataaaa
     6661 aaattttcat acttcttgct tttatttaaa ccattgaatg atttcttttg aacaaaacta
     6721 cctgtttcac caaaggaaat agaaagaaaa aatcaattag aagaaaacaa aa
//
