LOCUS       sequence-210306-        4143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CinRAM + PCin-YFP Reporter. OHC14 sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4143)
  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 4143)
  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.1642
FEATURES             Location/Qualifiers
     source          1..4143
                     /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(1910..1929)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2294..2839)
                     /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(2374..2393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2928..3743)
                     /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(2928..3743)
                     /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     3651..3670
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3735..3754
                     /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 ttgagaatac ctatagcgaa aagttcgagt ccgcgttcga acagatcaaa
     1021 gcggcggcca acgtggatgc cgccatccgt attctccagg cggaatataa cctcgatttc
     1081 gtcacctacc atctcgccca gacaatcgcg agcaagatcg attcgccctt cgtgcgcacc
     1141 acctatccgg atgcctgggt ttcccgttac ctcctcaact gctatgtgaa ggtcgatccg
     1201 atcatcaagc agggcttcga acgccagctg cccttcgact ggagcgaggt cgaaccgacg
     1261 ccggaggcct atgccatgct ggtcgacgcc cagaaacacg gcatcgatga caatggctac
     1321 tccatccccg tcgccgacaa ggcgcagcgc cgcgccctgc tgtcgctgaa tgcccatata
     1381 ccggccgacg aatggaccga gctcgtgcgc cgctgccgca atgagtggat cgagatcgcc
     1441 catctgatcc accgcaaggc cgtatatgag ctgcatggcg aaaacgatcc ggtgccggca
     1501 ttgtcgccgc gcgagatcga gtgtctgcac tggaccgccc tcggcaagga ttacaaggat
     1561 atttcggtca tcctgggcat atcagagcat accacacgcg attacctgaa aaccgcccgc
     1621 ttcaggctcg gctgcaccac gatctcggcc gccgcgtcgc gggctgttca attgcgcatc
     1681 atcaatccct ataggatccg catgacgcga cgtaattggt aatagcctaa ttggtaacga
     1741 atcagacaat tgacggctcg agggagtagc atagggtttg cagaatccct gcttcgtcca
     1801 tttgacaggc acattatgca tcgatgataa gctgtcaaac atgagcagat cctctacgcc
     1861 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     1921 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcaa atattttatc
     1981 tgaggtgctt cctcgctcac tgactcgctg cacgaggcag acctcagcgc tagcggagtg
     2041 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     2101 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     2161 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     2221 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     2281 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     2341 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
     2401 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     2461 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     2521 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     2581 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     2641 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     2701 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     2761 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     2821 cagaccaaaa cgatctcaag aagatcatct tattaagggg tctgacgctc agtggaacga
     2881 aaaatcaatc taaagtatat atgagtaaac ttggtctgac agttacctta gaaaaactca
     2941 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     3001 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     3061 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     3121 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     3181 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     3241 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     3301 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     3361 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     3421 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     3481 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     3541 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     3601 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     3661 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct ggagcaagac
     3721 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     3781 tttattgttc atgatgatat atttttatct tgtgcaatgt acatcagaga ttttgagaca
     3841 caaccaatta ttgaaggcct ccctaacggg gggccttttt ttgtttctgg tctcccgctt
     3901 aacgatcgtt ggctgccctt tgtgcgtcca aacggacgca cggcgctcta aagcgggtcg
     3961 cgatctttca gattcgctcc tcgcgctttc agtctttgtt ttggcgcatg tcgttatcgc
     4021 aaaaccgctg cacacttttg cgcgacatgc tctgatcccc ctcatctggg ggggcctatc
     4081 tgagggaatt tccgatccgg ctcgcctgaa ccattctgct ttccacgaac ttgaaaacgc
     4141 tag
//
