LOCUS       sequence-210282-        4558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MphRAM + EryR + PMph-YFP Reporter. Ery sensor for use in 
            non-Marionette strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4558)
  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 4558)
  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.969
FEATURES             Location/Qualifiers
     source          1..4558
                     /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(2499..2518)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2883..3428)
                     /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(2963..2982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3517..4332)
                     /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(3517..4332)
                     /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     4240..4259
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4324..4343
                     /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 atttagggag
      961 gtgaatatgc ctcgtccgaa actgaaaagt gatgatgaag ttctggaagc agcaaccgtt
     1021 gttctgaaac gttgtggtcc gattgaattt accctgagcg gtgttgcaaa agaagttggt
     1081 ctgagtcgcg cagcactgat tcagcgtttt accaatcgtg ataccctgct ggttcgtatg
     1141 atggaacgtg gtgttgaaca ggttcgtcat tatctgaatg caattccgat tggtgcaggt
     1201 ccgcagggtc tgtgggaatt tctgcaggtt ctggttcgta gcatggatac ccgtaatgat
     1261 ttcagcgtga actatctgat cagctggtat gaactgcagg ttccggaact gcgtaccctg
     1321 gcaattcagc gtaatcgtgc agttgttgaa ggtattcgta aacgtctgcc tccgggtgca
     1381 ccggcagcag cagaactgct gctgcatagc gttattgccg gtgcaaccat gcagtgggca
     1441 gttgatccgg atggtgaact ggcagatcat gttctggcac agattgcagc aattctgtgt
     1501 ctgatgtttc cggaacatga tgattttcag ctgctgcagg cacatgcata ataatattga
     1561 aaaaggaaga gtatgaacga gaaaaatata aaacacagtc aaaactttat tacttcaaaa
     1621 cataatatag ataaaataat gacaaatata agattaaatg aacatgataa tatctttgaa
     1681 atcggctcag gaaaaggcca ttttaccctt gaattagtaa agaggtgtaa tttcgtaact
     1741 gccattgaaa tagaccataa attatgcaaa actacagaaa ataaacttgt tgatcacgat
     1801 aatttccaag ttttaaacaa ggatatattg cagtttaaat ttcctaaaaa ccaatcctat
     1861 aaaatatatg gtaatatacc ttataacata agtacggata taatacgcaa aattgttttt
     1921 gatagtatag ctaatgagat ttatttaatc gtggaatacg ggtttgctaa aagattatta
     1981 aatacaaaac gctcattggc attactttta atggcagaag ttgatatttc tatattaagt
     2041 atggttccaa gagaatattt tcatcctaaa cctaaagtga atagctcact tatcagatta
     2101 agtagaaaaa aatcaagaat atcacacaaa gataaacaaa agtataatta tttcgttatg
     2161 aaatgggtta acaaagaata caagaaaata tttacaaaaa atcaatttaa caattcctta
     2221 aaacatgcag gaattgacga tttaaacaat attagctttg aacaattctt atctcttttc
     2281 aatagctata aattatttaa taagtgataa ggatcctaat tggtaacgaa tcagacaatt
     2341 gacggctcga gggagtagca tagggtttgc agaatccctg cttcgtccat ttgacaggca
     2401 cattatgcat cgatgataag ctgtcaaaca tgagcagatc ctctacgccg gacgcatcgt
     2461 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     2521 tggggaagat cgggctcgcc acttcgggct catgagcaaa tattttatct gaggtgcttc
     2581 ctcgctcact gactcgctgc acgaggcaga cctcagcgct agcggagtgt atactggctt
     2641 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     2701 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     2761 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     2821 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     2881 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     2941 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     3001 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     3061 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     3121 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3181 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     3241 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     3301 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     3361 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     3421 gatctcaaga agatcatctt attaaggggt ctgacgctca gtggaacgaa aaatcaatct
     3481 aaagtatata tgagtaaact tggtctgaca gttaccttag aaaaactcat cgagcatcaa
     3541 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     3601 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     3661 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     3721 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     3781 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     3841 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     3901 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     3961 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     4021 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     4081 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     4141 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     4201 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     4261 atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg
     4321 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     4381 tgatgatata tttttatctt gtgcaatgta catcagagat tttgagacac aaccaattat
     4441 tgaaggcctc cctaacgggg ggcctttttt tgtttctggt ctcccgctta acgatcgttg
     4501 gctgggattg aatataaccg acgtgactgt tacatttagg tggctaaacc cgtcaaag
//
