LOCUS       sequence-210311-        4098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PcaUAM + P3B5B-YFP Reporter. DHBA sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4098)
  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 4098)
  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.690
FEATURES             Location/Qualifiers
     source          1..4098
                     /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(1998..2017)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2382..2927)
                     /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(2462..2481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3016..3831)
                     /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(3016..3831)
                     /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     3739..3758
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3823..3842
                     /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 gtgaatatgt ggtcgaacat ggatgacaag aaagtgaaag aggagaatat tctgcacaat
     1021 tccaccaaca agaagatcat ccgccacgaa gattttgtag ccggcattag caaagggatg
     1081 gcgattctgg attcgtttgg tacagatcgt catcgcctca atatcaccat ggccgcagag
     1141 aaaaccggta tgacacgtgc agcagctcgt cgccacctgc ttactctgga gtatctgggc
     1201 tatctggaaa gtgacggcca ctacttctac ttaactccca aaatcctgaa attcagtggt
     1261 tcatatttgg gtggtgctca attgccgaaa atttcccaac cactgttgaa cttgcttacg
     1321 acccagacca gcctgattta cagcgtgatg gtgttggatg gctatgaagc cattaccatt
     1381 gcgcgttctg ccgctcatca gcaaaccgac cgcgttaacc cgtatggttt acatctcggg
     1441 aatcgcttac cagcgcatac aacgtcagcg ggcaaaatcc tgttagcgta tttggatgac
     1501 catgcccagc aagagtggct caatcagtac cctctgcaac ggctcacgaa atacacgtat
     1561 accaaccaca tcgactttct gcgccttttg agtgaaatca aggaacaggg ttggtgctat
     1621 agttcggaag aacacgaact gggagtacac gcccttgcgg ttccgattta cggacagcag
     1681 tctcgcgtcg tagcggcact gaacattgtc agcccgacaa tgcggaccac gaaagaatac
     1741 ctgattcagc atattctgcc gttactgcaa gaaactgcgc gtgaattgcg caatatcctg
     1801 taatgataag gatcctaatt ggtaacgaat cagacaattg acggctcgag ggagtagcat
     1861 agggtttgca gaatccctgc ttcgtccatt tgacaggcac attatgcatc gatgataagc
     1921 tgtcaaacat gagcagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     1981 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     2041 cttcgggctc atgagcaaat attttatctg aggtgcttcc tcgctcactg actcgctgca
     2101 cgaggcagac ctcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
     2161 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
     2221 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     2281 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     2341 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     2401 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     2461 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     2521 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     2581 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     2641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     2701 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     2761 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     2821 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     2881 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     2941 ttaaggggtc tgacgctcag tggaacgaaa aatcaatcta aagtatatat gagtaaactt
     3001 ggtctgacag ttaccttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     3061 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     3121 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     3181 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     3241 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     3301 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     3361 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     3421 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     3481 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     3541 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     3601 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     3661 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     3721 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     3781 ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct
     3841 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     3901 tgcaatgtac atcagagatt ttgagacaca accaattatt gaaggcctcc ctaacggggg
     3961 gccttttttt gtttctggtc tcccgcttaa cgatcgttgg ctgttttgtt cgattatcga
     4021 acaaattatt gaaatatcga acaaaacctc taaactactg tggcactgaa tcaaaaaatt
     4081 ataaaccatg atcagaag
//
