LOCUS       sequence-210300-        3834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PhlFAM + PPhlF-YFP Reporter. DAPG sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3834)
  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 3834)
  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.847
FEATURES             Location/Qualifiers
     source          1..3834
                     /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(1761..1780)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2145..2690)
                     /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(2225..2244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2779..3594)
                     /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(2779..3594)
                     /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     3502..3521
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3586..3605
                     /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 attcatgggg
      961 gtgaatatgg cacgtacccc gagccgtagc agcattggta gcctgcgtag tccgcatacc
     1021 cataaagcaa ttctgaccag caccattgaa atcctgaaag aatgtggtta tagcggtctg
     1081 agcattgaaa gcgtggcacg tcgcgccggt gcaggcaaac cgaccattta tcgttggtgg
     1141 accaacaaag cagcactgat tgccgaagtg tatgaaaatg aaatcgaaca ggtacgtaaa
     1201 tttccggatt tgggtagctt taaagccgat ctggattttc tgctgcataa tctgtggaaa
     1261 gtttggcgtg aaaccatttg tggtgaagca tttcgttgtg ttattgcaga agcacagttg
     1321 gaccctgtaa ccctgaccca actgaaagat cagtttatgg aacgtcgtcg tgagataccg
     1381 aaaaaactgg ttgaagatgc cattagcaat ggtgaactgc cgaaagatat caatcgtgaa
     1441 ctgctgctgg atatgatttt tggtttttgt tggtatcgcc tgctgaccga acagttgacc
     1501 gttgaacagg atattgaaga atttaccttc ctgctgatta atggtgtttg tccgggtaca
     1561 cagtgttgat aaggatccta attggtaacg aatcagacaa ttgacggctc gagggagtag
     1621 catagggttt gcagaatccc tgcttcgtcc atttgacagg cacattatgc atcgatgata
     1681 agctgtcaaa catgagcaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     1741 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     1801 ccacttcggg ctcatgagca aatattttat ctgaggtgct tcctcgctca ctgactcgct
     1861 gcacgaggca gacctcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg
     1921 agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca
     1981 gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     2041 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     2101 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     2161 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     2221 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     2281 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     2341 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     2401 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     2461 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     2521 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     2581 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     2641 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     2701 ttattaaggg gtctgacgct cagtggaacg aaaaatcaat ctaaagtata tatgagtaaa
     2761 cttggtctga cagttacctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2821 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2881 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     2941 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3001 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     3061 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3121 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     3181 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3241 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     3301 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3361 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3421 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     3481 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3541 gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc
     3601 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3661 ttgtgcaatg tacatcagag attttgagac acaaccaatt attgaaggcc tccctaacgg
     3721 ggggcctttt tttgtttctg gtctcccgct taacgatcgt tggctgcgac gtacggtgga
     3781 atctgattcg ttaccaattg acatgatacg aaacgtaccg tatcgttaag gtag
//
