LOCUS       sequence-267011-        7366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pREV1_ NLS_FdeR-N_2NLS-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7366)
  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 7366)
  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_009_026_096
FEATURES             Location/Qualifiers
     source          1..7366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             971..991
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             995..1015
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2884..3687
                     /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"
     CDS             complement(4329..5144)
                     /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(4329..5144)
                     /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     5052..5071
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5136..5155
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5369..5957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5858..5877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7346..7366
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggttctcgtt tcaacaagct cgacctcaat cttctggtcg ccctggatgc actgctcacg
       61 gagatgagca tcagccgcgc cgccgaaaag atccatctga gccagtcggc catgagcaat
      121 gccctggcgc ggctgcgcga gtatttcgat gatgaattgc tgatccaggt gggccggcgc
      181 atggagccca cgccgcgcgc cgaggtgctc aaggatgcgg tgcatgatgt gctgcggcgt
      241 atcgatggct ccatcgcggc gctgccggcc ttcgtgccgg ccgagtccac gcgcgagttt
      301 cgcatctcgg tttcggactt tacgctctcc gtcctcatcc cccgggtgct ggcgcgcgcg
      361 cacgccgagg gcaagcacat ccgctttgcc ctgatgccgc aggtgcaaga cccgacccgc
      421 tcgctggatc gggccgaggt ggacctgctg gtcttgccgc aggaattctg cacgcccgat
      481 catcctgccg aagaggtctt ccgcgaacgg catgtctgcg tggtctggcg cgacagtgcg
      541 ctggcgcaag gcgagctgac gctggaacgc tacatggcct caggccatgt ggtgatggtg
      601 ccgcctgggg ccaatgcgtc gtcggtggag gcgtggatgg ccaggaagct gggctttgcg
      661 cgccgggtgg aagtgaccag cttcagcttc gcttctgcgc tggcgctggt acaggggacg
      721 gaccgcatcg ccacggtgca tgcccggctg gcgcagctgc tggctccgca atggccggtg
      781 gtgatcaagg agagtccgct gtcgctgggc gagatgcggc agatgatgca gtggcatcgc
      841 taccgcagca atgatcctgg catccagtgg ctgcgtcggg tgtttctgga gagtgcgcag
      901 gagatggatg cggcgctgcc aggcatctgc ggatcctgag accaggctgc caggcatctg
      961 cggatccgat ccaaaaaaga agagaaaggt agatccaaaa aagaagagaa aggtataact
     1021 cgagtggcat tgaattgaat tgaaatcgat agatcaattt ttttcttttc tctttcccca
     1081 tcctttacgc taaaataata gtttatttta ttttttgaat attttttatt tatatacgta
     1141 tatatagact attatttatc ttttaatgat tattaagatt tttattaaaa aaaaattcgc
     1201 tcctctttta atgcctttat gcagtttttt tttcccattc gatatttcta tgtgctgcca
     1261 aaccagatgt caacacagct acaacgaagc ttcgtctttt cttgaagaaa agaggaagct
     1321 cgctaaatgg gattccactt tccgttccct gccagctgat ggaaaaaggt tagtggaacg
     1381 atgaagaata aaaagagaga tccactgagg tgaaatttca gctgacagcg agtttcatga
     1441 tcgtgatgaa caatggtaac gagttgtggc tgttgccagg gagggtggtt ctcaccaaat
     1501 cgctacttgg gtttgagatt ggtaaaattg attgtttgga tggcaatcat ctatggaatg
     1561 gatattatat aacaaagaag aaataatgaa ctgattctct tcctccttct tgtcctttct
     1621 taattctgtt gtaattacct tcctttgtaa ttttttttgt aattattctt cttaataatc
     1681 caaacaaaca cacatattac aataagatct atggtttcaa agggcgaaga agacaatatg
     1741 gctattatta aggaattcat gagattcaag gtccacatgg aaggttctgt caacggtcac
     1801 gaattcgaaa ttgaaggtga aggtgaaggt agaccatacg aaggtaccca aaccgctaag
     1861 ttgaaggtca ccaagggtgg tccattgcca ttcgcttggg atattttgtc tccacaattc
     1921 atgtacggtt ctaaggctta cgtcaagcac ccagctgata ttccagatta cttgaagttg
     1981 tctttcccag aaggtttcaa gtgggaaaga gtcatgaact tcgaagatgg tggtgtcgtc
     2041 accgtcaccc aagattcttc tttgcaagat ggtgaattca tttacaaggt caagttgaga
     2101 ggtaccaact tcccatctga tggtccagtc atgcaaaaga agaccatggg ttgggaagct
     2161 tcttctgaaa gaatgtaccc agaagatggt gctttgaagg gtgaaattaa gcaaagattg
     2221 aagttgaagg atggtggtca ctacgatgct gaagtcaaga ccacctacaa ggctaagaag
     2281 ccagtccaat tgccaggtgc ttacaacgtc aacattaagt tggatattac ctctcacaac
     2341 gaagattaca ccattgtcga acaatacgaa agagctgaag gtagacactc taccggtggt
     2401 atggatgaat tgtacaagta aatcctaact cgagataaag caatcttgat gaggataatg
     2461 attttttttt gaatatacat aaatactacc gtttttctgc tagattttgt gatgacgtaa
     2521 ataagtacat attacttttt aagccaagac aagattaagc attaacttta cccttttctt
     2581 tctaagtttc aatattagtt atcactgttt aaaagttatg gcgagaacgt cggcggttaa
     2641 aatatattac cctgaacggc tgagcagtta cagagatgtt acgaaccact agtgcactgc
     2701 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     2761 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     2821 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     2881 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     2941 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     3001 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     3061 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     3121 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     3181 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     3241 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     3301 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     3361 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     3421 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     3481 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     3541 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     3601 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     3661 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     3721 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta gagcacttga
     3781 atccactgcc ccgggaatct cggtcgtaat gatttctata atgacgaaaa aaaaaaaatt
     3841 ggaaagaaaa agcttcatgg cctttataaa aaggaactat ccaatacctc gccagaacca
     3901 agtaacagta ttttacgggg cacaaatcaa gaacaataag acaggactgt aaagatggac
     3961 gcattgaact ccaaagaaca acaagagttc caaaaagtag tggaacaaaa gcaaatgaag
     4021 gatttcatgc gtttgtactc taatctggta gaaagatgtt tcacagactg tgtcaatgac
     4081 ttcacaacat caaagctaac caataaggaa caaacatgca tcatgaagtg ctcagaaaag
     4141 ttcttgaagc atagcgaacg tgtagggcag cgtttccaag aacaaaacgc tgccttggga
     4201 caaggcttgg gccgataagg tgtactggcg tatatatatc taattatgta tctctggtgt
     4261 agcccatttt tagcatgtaa atataaagac cgagcggccg cgtgttacaa ccaattaacc
     4321 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     4381 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     4441 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     4501 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     4561 gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca
     4621 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     4681 cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca attacaaaca
     4741 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     4801 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     4861 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     4921 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     4981 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     5041 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     5101 aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     5161 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     5221 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     5281 ttgaaggatc agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5341 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5401 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5461 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5521 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5581 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5641 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5701 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5761 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5821 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5881 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5941 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6001 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg
     6061 ccgccaatgc taaattcgag tgaaacacag gaagatcaga aaatcctcat ttcatccata
     6121 ttaacaataa tttcaaatgt ttatttgcat tatttgaaac taggcaagac aagcaacgaa
     6181 acgtttttga aaattttgag tattttcaat aaatttgtag aggactcaga tattgaaaaa
     6241 aagctacagc aattaatact tgataagaag agtattgaga agggcaacgg ttcatcatct
     6301 catggatctg cacatgaaca aacaccagag tcaaacgacg ttgaaattga ggctactgcg
     6361 ccaattgatg acaatacaga cgatgataac aaaccgaagt tatctgatgt agaaaaggat
     6421 taaagatgct aagagatagt gatgatattt cataaataat gtaattctat atatgttaat
     6481 tacctttttt gcgaggcata tttatggtga aggataagtt ttgaccatca aagaaggtta
     6541 atgtggctgt ggtttcaggg tccataaagc ccacatggat aacattaccc ctgaattcgc
     6601 atctagactg aactggccga taattgcaga cgaacggtgt tgttatccga tacaaccgga
     6661 tatttttctt ttaatgagtc taaaccgtga tagcttcagg ttaatacaat caaaaaaagc
     6721 tcaaatattc ttttaatgcc gcgttcacag attccaattg aatacaacta ggtagttcat
     6781 tatatgaagc ctttgctact atttttcact atagtctgcc ttcaccttaa tgcagacatc
     6841 cacatatttt aatcacttta aaataaaaag gaagatatat tagaagctat gatccaatct
     6901 gtaagccaga ttaaaattca cgaactcttc tttcatttga attgaatgct ttgagttggg
     6961 gtagattatc gcaaattact catcacattt attgactacg aacttgctga tgtccttttt
     7021 ttatttatat ttttcttcag tgaagcgatt ttttttttac acagaccaag acggaaaaaa
     7081 gtagctaagg aagaaaacaa aatcatgaaa aaaatgtgaa gtgatcatgc acatcgcatc
     7141 aacttaaaca ttggcttaga gatatataga gttagagttt acggcaacct ttaagcacca
     7201 ataccttttg gcatagtcta aagacctggt tcttaatttt aaacaaattt aactaaagat
     7261 ttccctatca aagaagtaac gagttgacag attttctcaa aataaatcga tactgcattt
     7321 ctaggcatat ccagcgagat ctatgccaaa gaagaagaga aaggtt
//
