LOCUS       sequence-213316-        3852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TtgRAM + PTtg-YFP Reporter. Nar sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3852)
  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 3852)
  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.661
FEATURES             Location/Qualifiers
     source          1..3852
                     /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(1794..1813)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2178..2723)
                     /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(2258..2277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2812..3627)
                     /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(2812..3627)
                     /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     3535..3554
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3619..3638
                     /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 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggag
      961 gtgaatatgg ttcgtcgtac caaagaagag gcacaagaaa cccgtgcaca gattattgaa
     1021 gcagcagaac gtgcattcta taaacgtggt gttgcacgta ccaccctggc agatattgca
     1081 gaactggcag gcgttacccg tggtgcaatt tattggcatt ttaacaacaa agccgaactg
     1141 gttcaggcac tgctggatag cctgcatgaa acccatgatc atctggcacg tgcaagcgaa
     1201 agcgaagatg aagttgatcc gctgggttgt atgcgtaaac tgctgctgca ggtttttaat
     1261 gaactggttc tggatgcacg tacccgtcgt attaatgaaa tcctgcatca caaatgcgag
     1321 ttcaccgatg atatgtgtga aattcgtcag cagcatcaga gcgcagttct ggattgtcat
     1381 aaaggtatta ccctgacact ggcaaatgta gttcgtcgtg gtcagctgcc tggtgaactg
     1441 gatgcagaac gtgccgcagt tgcaatgttt gcctatgttg atggtctgat tcgtcgttgg
     1501 ctgctgctgc cggatagcgt tgatctgctg ggtgatgttg aaaaatgggt tgataccggt
     1561 ctggatatgc tgcgtctgag tccggcactg cgtaaataat gataaggatc ctaattggta
     1621 acgaatcaga caattgacgg ctcgagggag tagcataggg tttgcagaat ccctgcttcg
     1681 tccatttgac aggcacatta tgcatcgatg ataagctgtc aaacatgagc agatcctcta
     1741 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     1801 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcaaatattt
     1861 tatctgaggt gcttcctcgc tcactgactc gctgcacgag gcagacctca gcgctagcgg
     1921 agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg
     1981 caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc
     2041 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     2101 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     2161 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     2221 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2281 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     2341 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     2401 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     2461 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     2521 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     2581 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     2641 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta
     2701 cgcgcagacc aaaacgatct caagaagatc atcttattaa ggggtctgac gctcagtgga
     2761 acgaaaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac cttagaaaaa
     2821 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     2881 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     2941 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3001 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3061 tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3121 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3181 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3241 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3301 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3361 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3421 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3481 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     3541 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctggagca
     3601 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     3661 cagttttatt gttcatgatg atatattttt atcttgtgca atgtacatca gagattttga
     3721 gacacaacca attattgaag gcctccctaa cggggggcct ttttttgttt ctggtctccc
     3781 gcttaacgat cgttggctgc acccagcagt atttacaaac aaccatgaat gtaagtatat
     3841 tccttagcaa ag
//
