LOCUS       sequence-266973-        6748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pRPL18B_NLS_FdeR-N_tPGK1-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6748)
  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 6748)
  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_002_026_MV1
FEATURES             Location/Qualifiers
     source          1..6748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             269..289
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3091..3894
                     /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    3977..4480
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4524..5339)
                     /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(4524..5339)
                     /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     5247..5266
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5331..5350
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5564..6152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6053..6072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcaattgac acttcattag ccatcacaca aaactctttc ttgctggagc ttcttttaaa
       61 aaagacctca gtacaccaaa cacgttaccc gacctcgtta ttttacgaca actatgataa
      121 aattctgaag aaaaaataaa aaaattttca tacttcttgc ttttatttaa accattgaat
      181 gatttctttt gaacaaaact acctgtttca ccaaaggaaa tagaaagaaa aaatcaatta
      241 gaagaaaaca aaaaacaaaa gatctatgcc aaagaagaag agaaaggttg gttctcgttt
      301 caacaagctc gacctcaatc ttctggtcgc cctggatgca ctgctcacgg agatgagcat
      361 cagccgcgcc gccgaaaaga tccatctgag ccagtcggcc atgagcaatg ccctggcgcg
      421 gctgcgcgag tatttcgatg atgaattgct gatccaggtg ggccggcgca tggagcccac
      481 gccgcgcgcc gaggtgctca aggatgcggt gcatgatgtg ctgcggcgta tcgatggctc
      541 catcgcggcg ctgccggcct tcgtgccggc cgagtccacg cgcgagtttc gcatctcggt
      601 ttcggacttt acgctctccg tcctcatccc ccgggtgctg gcgcgcgcgc acgccgaggg
      661 caagcacatc cgctttgccc tgatgccgca ggtgcaagac ccgacccgct cgctggatcg
      721 ggccgaggtg gacctgctgg tcttgccgca ggaattctgc acgcccgatc atcctgccga
      781 agaggtcttc cgcgaacggc atgtctgcgt ggtctggcgc gacagtgcgc tggcgcaagg
      841 cgagctgacg ctggaacgct acatggcctc aggccatgtg gtgatggtgc cgcctggggc
      901 caatgcgtcg tcggtggagg cgtggatggc caggaagctg ggctttgcgc gccgggtgga
      961 agtgaccagc ttcagcttcg cttctgcgct ggcgctggta caggggacgg accgcatcgc
     1021 cacggtgcat gcccggctgg cgcagctgct ggctccgcaa tggccggtgg tgatcaagga
     1081 gagtccgctg tcgctgggcg agatgcggca gatgatgcag tggcatcgct accgcagcaa
     1141 tgatcctggc atccagtggc tgcgtcgggt gtttctggag agtgcgcagg agatggatgc
     1201 ggcgctgcca ggcatctgct gaatcctaac tcgagattga attgaattga aatcgataga
     1261 tcaatttttt tcttttctct ttccccatcc tttacgctaa aataatagtt tattttattt
     1321 tttgaatatt ttttatttat atacgtatat atagactatt atttatcttt taatgattat
     1381 taagattttt attaaaaaaa aattcgctcc tcttttaatg cctttatgca gttttttttt
     1441 cccattcgat atttctatgt gctgccaaac cagatgtcaa cacagctaca acgaagcttc
     1501 gtcttttctt gaagaaaaga ggaagctcgc taaatgggat tccactttcc gttccctgcc
     1561 agctgatgga aaaaggttag tggaacgatg aagaataaaa agagagatcc actgaggtga
     1621 aatttcagct gacagcgagt ttcatgatcg tgatgaacaa tggtaacgag ttgtggctgt
     1681 tgccagggag ggtggttctc accaaatcgc tacttgggtt tgagattggt aaaattgatt
     1741 gtttggatgg caatcatcta tggaatggat attatataac aaagaagaaa taatgaactg
     1801 attctcttcc tccttcttgt cctttcttaa ttctgttgta attaccttcc tttgtaattt
     1861 tttttgtaat tattcttctt aataatccaa acaaacacac atattacaat aagatctatg
     1921 gtttcaaagg gcgaagaaga caatatggct attattaagg aattcatgag attcaaggtc
     1981 cacatggaag gttctgtcaa cggtcacgaa ttcgaaattg aaggtgaagg tgaaggtaga
     2041 ccatacgaag gtacccaaac cgctaagttg aaggtcacca agggtggtcc attgccattc
     2101 gcttgggata ttttgtctcc acaattcatg tacggttcta aggcttacgt caagcaccca
     2161 gctgatattc cagattactt gaagttgtct ttcccagaag gtttcaagtg ggaaagagtc
     2221 atgaacttcg aagatggtgg tgtcgtcacc gtcacccaag attcttcttt gcaagatggt
     2281 gaattcattt acaaggtcaa gttgagaggt accaacttcc catctgatgg tccagtcatg
     2341 caaaagaaga ccatgggttg ggaagcttct tctgaaagaa tgtacccaga agatggtgct
     2401 ttgaagggtg aaattaagca aagattgaag ttgaaggatg gtggtcacta cgatgctgaa
     2461 gtcaagacca cctacaaggc taagaagcca gtccaattgc caggtgctta caacgtcaac
     2521 attaagttgg atattacctc tcacaacgaa gattacacca ttgtcgaaca atacgaaaga
     2581 gctgaaggta gacactctac cggtggtatg gatgaattgt acaagtaaat cctaactcga
     2641 gataaagcaa tcttgatgag gataatgatt tttttttgaa tatacataaa tactaccgtt
     2701 tttctgctag attttgtgat gacgtaaata agtacatatt actttttaag ccaagacaag
     2761 attaagcatt aactttaccc ttttctttct aagtttcaat attagttatc actgtttaaa
     2821 agttatggcg agaacgtcgg cggttaaaat atattaccct gaacggctga gcagttacag
     2881 agatgttacg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     2941 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     3001 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     3061 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     3121 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     3181 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     3241 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     3301 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     3361 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     3421 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     3481 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     3541 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     3601 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     3661 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     3721 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     3781 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     3841 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     3901 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     3961 cagttattac ccgagtatca cgtgctataa aaataattat aatttaaatt ttttaatata
     4021 aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt
     4081 tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt atcttgctct
     4141 tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa aaccacacac
     4201 gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct atttaatctg
     4261 cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct
     4321 ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta ctttctaaaa
     4381 tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt attccatcat
     4441 taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtcccgagc ggccgcgtgt
     4501 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
     4561 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     4621 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     4681 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     4741 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct
     4801 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     4861 aaaccgttat tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa
     4921 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     4981 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     5041 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     5101 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     5161 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     5221 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     5281 tccatgttgg aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata
     5341 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     5401 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat
     5461 cgaacttttg ctgagttgaa ggatcagtca tgaccaaaat cccttaacgt gagttttcgt
     5521 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5581 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5641 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5701 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5761 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5821 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5881 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5941 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6001 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6061 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6121 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6181 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6241 tggataaccg tgcggccgcc cctgaattcg catctagact gaactggccg ataattgcag
     6301 acgaacgaag aggatgtcca atattttttt taaggaataa ggatacttca agactagatt
     6361 cccccctgca ttcccatcag aaccgtaaac cttggcgctt tccttgggaa gtattcaaga
     6421 agtgccttgt ccggtttctg tggctcacaa accagcgcgc ccgatatggc tttcttttca
     6481 cttatgaatg taccagtacg ggacaattag aacgctcctg taacaatctc tttgcaaatg
     6541 tggggttaca ttctaaccat gtcacactgc tgacgaaatt caaagtaaaa aaaaatggga
     6601 ccacgtcttg agaacgatag attttcttta ttttacattg aacagtcgtt gtctcagcgc
     6661 gctttatgtt ttcattcata cttcatatta taaaataaca aaagaagaat ttcatattca
     6721 cgcccaagaa atcaggctgc tttccaaa
//
