LOCUS       sequence-210309-        3951 bp    DNA     circular SYN 14-APR-2025
DEFINITION  VanRAM + PVanCC-YFP Reporter. Van sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3951)
  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 3951)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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.773
FEATURES             Location/Qualifiers
     source          1..3951
                     /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     complement(1894..1913)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2278..2823)
                     /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(2358..2377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2912..3727)
                     /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(2912..3727)
                     /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     3635..3654
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3719..3738
                     /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 tactggagtt cgtgaccgcc
      781 gccgggatca ctctcggcat ggacgagctg tacaagtaac tcggtaccaa attccagaaa
      841 agaggcctcc cgaaaggggg gccttttttc gttttggtcc aatggcggcg cgccatcgaa
      901 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcaggggg
      961 gtgaataatg gacatgcctc gtattaaacc gggtcagcgt gttatgatgg cactgcgtaa
     1021 aatgattgca agcggtgaaa tcaaaagtgg tgaacgtatt gcagaaattc cgaccgcagc
     1081 agcactgggt gttagccgta tgccggttcg tatcgcactg cgttcactgg aacaagaagg
     1141 tctggttgtt cgtctgggtg cacgtggtta tgcagcccgt ggtgttagca gcgatcagat
     1201 tcgtgatgca attgaagttc gtggtgttct ggaaggtttt gcagcacgtc gtctggcaga
     1261 acgtggtatg accgcagaaa cccatgcacg ttttgttgta ctgattgcag aaggtgaagc
     1321 actgtttgca gccggtcgcc tgaatggtga agatctggat cgttatgccg catataatca
     1381 ggcatttcat gataccctgg ttagcgcagc aggtaatggt gcagttgaaa gcgcactggc
     1441 acgtaatggt tttgaaccgt ttgcagcagc cggtgcactg gccctggatc tgatggacct
     1501 gtctgccgaa tatgaacatc tgctggcagc acatcgtcag catcaggcag ttctggatgc
     1561 agttagctgt ggtgatgccg aaggtgcaga acgtattatg cgtgatcatg cactggcagc
     1621 aattcgtaat gcaaaagttt ttgaagcagc agcaagcgca ggcgcaccgc tgggtgcagc
     1681 atggtcaatt cgtgcagatt gataaggatc ctaattggta acgaatcaga caattgacgg
     1741 ctcgagggag tagcataggg tttgcagaat ccctgcttcg tccatttgac aggcacatta
     1801 tgcatcgatg ataagctgtc aaacatgagc agatcctcta cgccagacgc atcgtggccg
     1861 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     1921 aagatcgggc tcgccacttc gggctcatga gcaaatattt tatctgaggt gcttcctcgc
     1981 tcactgactc gctgcacgag gcagacctca gcgctagcgg agtgtatact ggcttactat
     2041 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     2101 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     2161 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     2221 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     2281 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     2341 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     2401 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     2461 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     2521 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2581 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     2641 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     2701 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     2761 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     2821 caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaaatc aatctaaagt
     2881 atatatgagt aaacttggtc tgacagttac cttagaaaaa ctcatcgagc atcaaatgaa
     2941 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     3001 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     3061 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     3121 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     3181 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     3241 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     3301 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     3361 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     3421 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     3481 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     3541 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     3601 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     3661 aatcagcatc catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat
     3721 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     3781 atatattttt atcttgtgca atgtacatca gagattttga gacacaacca attattgaag
     3841 gcctccctaa cggggggcct ttttttgttt ctggtctccc gcttaacgat cgttggctga
     3901 ttggatccaa ttgacagcta gctcagtcct aggtaccatt ggatccaata g
//
