LOCUS       sequence-266882-        6748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV1-pREV1_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_003_026_MV1
FEATURES             Location/Qualifiers
     source          1..6748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             66..86
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2888..3691
                     /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    3774..4277
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4321..5136)
                     /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(4321..5136)
                     /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     5044..5063
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5128..5147
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5361..5949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5850..5869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagttgacag attttctcaa aataaatcga tactgcattt ctaggcatat ccagcgagat
       61 ctatgccaaa gaagaagaga aaggttggtt ctcgtttcaa caagctcgac ctcaatcttc
      121 tggtcgccct ggatgcactg ctcacggaga tgagcatcag ccgcgccgcc gaaaagatcc
      181 atctgagcca gtcggccatg agcaatgccc tggcgcggct gcgcgagtat ttcgatgatg
      241 aattgctgat ccaggtgggc cggcgcatgg agcccacgcc gcgcgccgag gtgctcaagg
      301 atgcggtgca tgatgtgctg cggcgtatcg atggctccat cgcggcgctg ccggccttcg
      361 tgccggccga gtccacgcgc gagtttcgca tctcggtttc ggactttacg ctctccgtcc
      421 tcatcccccg ggtgctggcg cgcgcgcacg ccgagggcaa gcacatccgc tttgccctga
      481 tgccgcaggt gcaagacccg acccgctcgc tggatcgggc cgaggtggac ctgctggtct
      541 tgccgcagga attctgcacg cccgatcatc ctgccgaaga ggtcttccgc gaacggcatg
      601 tctgcgtggt ctggcgcgac agtgcgctgg cgcaaggcga gctgacgctg gaacgctaca
      661 tggcctcagg ccatgtggtg atggtgccgc ctggggccaa tgcgtcgtcg gtggaggcgt
      721 ggatggccag gaagctgggc tttgcgcgcc gggtggaagt gaccagcttc agcttcgctt
      781 ctgcgctggc gctggtacag gggacggacc gcatcgccac ggtgcatgcc cggctggcgc
      841 agctgctggc tccgcaatgg ccggtggtga tcaaggagag tccgctgtcg ctgggcgaga
      901 tgcggcagat gatgcagtgg catcgctacc gcagcaatga tcctggcatc cagtggctgc
      961 gtcgggtgtt tctggagagt gcgcaggaga tggatgcggc gctgccaggc atctgctgaa
     1021 tcctaactcg agattgaatt gaattgaaat cgatagatca atttttttct tttctctttc
     1081 cccatccttt acgctaaaat aatagtttat tttatttttt gaatattttt tatttatata
     1141 cgtatatata gactattatt tatcttttaa tgattattaa gatttttatt aaaaaaaaat
     1201 tcgctcctct tttaatgcct ttatgcagtt tttttttccc attcgatatt tctatgtgct
     1261 gccaaaccag atgtcaacac agctacaacg aagcttcgtc ttttcttgaa gaaaagagga
     1321 agctcgctaa atgggattcc actttccgtt ccctgccagc tgatggaaaa aggttagtgg
     1381 aacgatgaag aataaaaaga gagatccact gaggtgaaat ttcagctgac agcgagtttc
     1441 atgatcgtga tgaacaatgg taacgagttg tggctgttgc cagggagggt ggttctcacc
     1501 aaatcgctac ttgggtttga gattggtaaa attgattgtt tggatggcaa tcatctatgg
     1561 aatggatatt atataacaaa gaagaaataa tgaactgatt ctcttcctcc ttcttgtcct
     1621 ttcttaattc tgttgtaatt accttccttt gtaatttttt ttgtaattat tcttcttaat
     1681 aatccaaaca aacacacata ttacaataag atctatggtt tcaaagggcg aagaagacaa
     1741 tatggctatt attaaggaat tcatgagatt caaggtccac atggaaggtt ctgtcaacgg
     1801 tcacgaattc gaaattgaag gtgaaggtga aggtagacca tacgaaggta cccaaaccgc
     1861 taagttgaag gtcaccaagg gtggtccatt gccattcgct tgggatattt tgtctccaca
     1921 attcatgtac ggttctaagg cttacgtcaa gcacccagct gatattccag attacttgaa
     1981 gttgtctttc ccagaaggtt tcaagtggga aagagtcatg aacttcgaag atggtggtgt
     2041 cgtcaccgtc acccaagatt cttctttgca agatggtgaa ttcatttaca aggtcaagtt
     2101 gagaggtacc aacttcccat ctgatggtcc agtcatgcaa aagaagacca tgggttggga
     2161 agcttcttct gaaagaatgt acccagaaga tggtgctttg aagggtgaaa ttaagcaaag
     2221 attgaagttg aaggatggtg gtcactacga tgctgaagtc aagaccacct acaaggctaa
     2281 gaagccagtc caattgccag gtgcttacaa cgtcaacatt aagttggata ttacctctca
     2341 caacgaagat tacaccattg tcgaacaata cgaaagagct gaaggtagac actctaccgg
     2401 tggtatggat gaattgtaca agtaaatcct aactcgagat aaagcaatct tgatgaggat
     2461 aatgattttt ttttgaatat acataaatac taccgttttt ctgctagatt ttgtgatgac
     2521 gtaaataagt acatattact ttttaagcca agacaagatt aagcattaac tttacccttt
     2581 tctttctaag tttcaatatt agttatcact gtttaaaagt tatggcgaga acgtcggcgg
     2641 ttaaaatata ttaccctgaa cggctgagca gttacagaga tgttacgaac cactagtgca
     2701 ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga
     2761 acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac
     2821 atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag
     2881 ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc
     2941 tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt
     3001 tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact
     3061 aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa
     3121 ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg
     3181 taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat
     3241 tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga
     3301 agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc
     3361 cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt
     3421 tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt
     3481 gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag
     3541 aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt
     3601 tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc
     3661 atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat
     3721 actaaactca caaattagag cttcaattta attatatcag ttattacccg agtatcacgt
     3781 gctataaaaa taattataat ttaaattttt taatataaat atataaatta aaaatagaaa
     3841 gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag
     3901 ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc
     3961 gaattgtttc atcttgtctg tgtagaaaac cacacacgaa aatcctgtga ttttacattt
     4021 tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat
     4081 atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt
     4141 ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata
     4201 aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa
     4261 gttacaggca agcgatccgt cccgagcggc cgcgtgttac aaccaattaa ccaattctga
     4321 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     4381 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     4441 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     4501 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     4561 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     4621 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     4681 cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     4741 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     4801 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     4861 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     4921 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     4981 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     5041 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     5101 cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     5161 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     5221 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
     5281 tcagtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5341 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5401 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5461 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5521 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5581 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5641 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5701 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5761 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5821 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5881 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5941 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6001 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgcccct
     6061 gaattcgcat ctagactgaa ctggccgata attgcagacg aacggtgttg ttatccgata
     6121 caaccggata tttttctttt aatgagtcta aaccgtgata gcttcaggtt aatacaatca
     6181 aaaaaagctc aaatattctt ttaatgccgc gttcacagat tccaattgaa tacaactagg
     6241 tagttcatta tatgaagcct ttgctactat ttttcactat agtctgcctt caccttaatg
     6301 cagacatcca catattttaa tcactttaaa ataaaaagga agatatatta gaagctatga
     6361 tccaatctgt aagccagatt aaaattcacg aactcttctt tcatttgaat tgaatgcttt
     6421 gagttggggt agattatcgc aaattactca tcacatttat tgactacgaa cttgctgatg
     6481 tccttttttt atttatattt ttcttcagtg aagcgatttt ttttttacac agaccaagac
     6541 ggaaaaaagt agctaaggaa gaaaacaaaa tcatgaaaaa aatgtgaagt gatcatgcac
     6601 atcgcatcaa cttaaacatt ggcttagaga tatatagagt tagagtttac ggcaaccttt
     6661 aagcaccaat accttttggc atagtctaaa gacctggttc ttaattttaa acaaatttaa
     6721 ctaaagattt ccctatcaaa gaagtaac
//
