LOCUS       sequence-210299-        3973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LuxR + PLuxB-YFP Reporter. OC6 sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3973)
  AUTHORS   Meyer AJ, Segall-Shapiro TH, Glassey E, Zhang J, Voigt CA
  TITLE     Escherichia coli ""Marionette"" strains with 12 highly optimized 
            small-molecule sensors.
  JOURNAL   Nat Chem Biol. 2018 Nov 26. pii: 10.1038/s41589-018-0168-3. doi: 
            10.1038/s41589-018-0168-3.
  PUBMED    30478458
REFERENCE   2  (bases 1 to 3973)
  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"; journalName: "Nat Chem 
            Biol. 2018 Nov 26. pii: 10.1038/s41589-018-0168-3. doi: 
            10.1038/s41589-018-0168-3."
COMMENT     Sequence Label: pAJM.474
FEATURES             Location/Qualifiers
     source          1..3973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..50
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(100..102,103..105,106..819)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(145..166)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(406..425)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     753..774
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1911..1930)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2295..2840)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2375..2394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2929..3744)
                     /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(2929..3744)
                     /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     3652..3671
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3736..3755
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa
       61 taattttgtt taatactaga gaaagagggg aaatactaga tggtgagcaa gggcgaggag
      121 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      181 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      241 atctgcacca caggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
      301 ggcctgcaat gcttcgcccg ctaccccgac cacatgaagc tgcacgactt cttcaagtcc
      361 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      421 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      481 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      541 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      601 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      661 ccaatcggcg acggccccgt gctgctgccc gacaaccact accttagcta ccagtccgcc
      721 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
      781 gccgggatca ctctcggcat ggacgagctg tacaagtaac tcggtaccaa attccagaaa
      841 agaggcctcc cgaaaggggg gccttttttc gttttggtcc aatggcggcg cgccatcgaa
      901 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
      961 gtgaatatga aaaacataaa tgccgacgac acatacagaa taattaataa aattaaagct
     1021 tgtagaagca ataatgatat taatcaatgc ttatctgata tgactaaaat ggtacattgt
     1081 gaatattatt tactcgcgat catttatcct cattctatgg ttaaatctga tatttcaatc
     1141 ctagataatt accctaaaaa atggaggcaa tattatgatg acgctaattt aataaaatat
     1201 gatcctatag tagattattc taactccaat cattcaccaa ttaattggaa tatatttgaa
     1261 aacaatgctg taaataaaaa atctccaaat gtaattaaag aagcgaaaac atcaggtctt
     1321 atcactgggt ttagtttccc tattcatacg gctaacaatg gcttcggaat gcttagtttt
     1381 gcacattcag aaaaagacaa ctatatagat agtttatttt tacatgcgtg tatgaacata
     1441 ccattaattg ttccttctct agttgataat tatcgaaaaa taaatatagc aaataataaa
     1501 tcaaacaacg atttaaccaa aagagaaaaa gaatgtttag cgtgggcatg cgaaggaaaa
     1561 agctcttggg atatttcaaa aatattaggt tgcagtgagc gtactgtcac tttccattta
     1621 accaatgcgc aaatgaaact caatacaaca aaccgctgcc aaagtatttc taaagcaatt
     1681 ttaacaggag caattgattg cccatacttt aaaaattgat aaggatccta attggtaacg
     1741 aatcagacaa ttgacggctc gagggagtag catagggttt gcagaatccc tgcttcgtcc
     1801 atttgacagg cacattatgc atcgatgata agctgtcaaa catgagcaga tcctctacgc
     1861 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     1921 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagca aatattttat
     1981 ctgaggtgct tcctcgctca ctgactcgct gcacgaggca gacctcagcg ctagcggagt
     2041 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
     2101 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
     2161 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
     2221 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
     2281 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
     2341 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc
     2401 ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta
     2461 tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa
     2521 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta
     2581 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa
     2641 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa
     2701 gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga
     2761 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc
     2821 gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct cagtggaacg
     2881 aaaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttacctt agaaaaactc
     2941 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     3001 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     3061 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     3121 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     3181 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     3241 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     3301 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     3361 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     3421 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     3481 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     3541 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     3601 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     3661 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga
     3721 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     3781 ttttattgtt catgatgata tatttttatc ttgtgcaatg tacatcagag attttgagac
     3841 acaaccaatt attgaaggcc tccctaacgg ggggcctttt tttgtttctg gtctcccgct
     3901 taacgatcgt tggctgacct gtaggatcgt acaggtttac gcaagaaaat ggtttgttac
     3961 agtcgaataa aag
//
