LOCUS       sequence-210295-        4184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NahRAM +PSalTTC-YFP Reporter. Sal sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4184)
  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 4184)
  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.771
FEATURES             Location/Qualifiers
     source          1..4184
                     /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(2061..2080)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2445..2990)
                     /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(2525..2544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3079..3894)
                     /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(3079..3894)
                     /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     3802..3821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3886..3905
                     /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 gcctttttcg ttttggtcca atggcggcgc gccatcgaat
      901 ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcatggggg
      961 ttgaatatgg aactgcgtga cctggattta aacctgctgg tggtgttcaa ccagttgctg
     1021 gtcgacagac gcgtctctgt cactgcggag aacctgggcc tgacccagcc tgccgtgagc
     1081 aatgcgctga aacgcctgcg cacctcgcta caggacccac tcttcgtgcg cacacatcag
     1141 ggaatggaac ccacacccta tgccgcgcat ctggccgagc acgtcacttc ggccatgcac
     1201 gcactgcgca acgccctaca gcaccatgaa agcttcgatc cgctgaccag cgagcgtacc
     1261 ttcaccctgg ccatgaccga cattggcgag atctacttca tgccgcggct gatggatgcg
     1321 ctggctcacc aggcccccaa ttgcgtgatc agtacggtgc gcgacagttc gatgagcctg
     1381 atgcaggcct tgcagaacgg aaccgtggac ttggccgtgg gcctgcttcc caatctgcaa
     1441 actggcttct ttcagcgccg gctgctccag aatcactacg tgtgcctatg tcgcaaggac
     1501 catccagtca cccgcgaacc cctgactctg gagcgcttct gttcctacgg ccacgtgcgt
     1561 gtcatcgccg ctggcaccgg ccacggcgag gtggacacgt acatgacacg ggtcggcatc
     1621 cggcgcgaca tccgtctgga agtgccgcac ttcgccgccg ttggccacat cctccagcgc
     1681 accgatctgc tcgccactgt gccgatatgt ttagccgact gctgcgtaga gcccttcggc
     1741 ctaagcgcct tgccgcaccc agtcgtcttg cctgaaatag ccatcaacat gttctggcat
     1801 gcgaagtacc acaaggacct agccaatatt tggttgcggc aactgatgtt tgacctgttt
     1861 acggattgat aaggatccta attggtaacg aatcagacaa ttgacggctc gagggagtag
     1921 catagggttt gcagaatccc tgcttcgtcc atttgacagg cacattatgc atcgatgata
     1981 agctgtcaaa catgagcaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     2041 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     2101 ccacttcggg ctcatgagca aatattttat ctgaggtgct tcctcgctca ctgactcgct
     2161 gcacgaggca gacctcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg
     2221 agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca
     2281 gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     2341 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     2401 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     2461 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     2521 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     2581 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     2641 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     2701 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     2761 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     2821 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     2881 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     2941 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     3001 ttattaaggg gtctgacgct cagtggaacg aaaaatcaat ctaaagtata tatgagtaaa
     3061 cttggtctga cagttacctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3121 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3181 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3241 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3301 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     3361 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3421 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     3481 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3541 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     3601 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3661 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3721 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     3781 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3841 gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc
     3901 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3961 ttgtgcaatg tacatcagag attttgagac acaaccaatt attgaaggcc tccctaacgg
     4021 ggggcctttt tttgtttctg gtctcccgct taacgatcgt tggctggggg cctcgcttgg
     4081 gttattgctg gtgcccggcc gggcgcaata ttcatgttga tgatttatta tatatcgagt
     4141 ggtgtattta tttatattgt ttgctccgtt accgttatta acag
//
