LOCUS       sequence-210312-        3816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BetIAM + PBetI-YFP Reporter. Cho sensor for use in non-Marionette 
            strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3816)
  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 3816)
  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.683
FEATURES             Location/Qualifiers
     source          1..3816
                     /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(1746..1765)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(2130..2675)
                     /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(2210..2229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2764..3579)
                     /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(2764..3579)
                     /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     3487..3506
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3571..3590
                     /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 attcacgggg
      961 gtgaatatgc cgaaactggg tatgcagagc attcgtcgtc gtcagctgat tgatgcaacc
     1021 ctggaagcaa ttaatgaagt tggtatgcat gatgcaacca ttgcacagat tgcacgtcgt
     1081 gccggtgtta gcaccggtat tattagccat tatttccgcg ataaaaacgg tctactggaa
     1141 gcaaccatgc gtgatattac cagccagctg cgtgatgcag ttctgaatcg tctgcatgca
     1201 ctgccgcagg gtagcgcaga acagcgtctg caggcaattg ttggtggtaa ttttgatgaa
     1261 acccaggtta gcagcgcagc aatgaaagca tggctggcat tttgggcaat cagcatgcat
     1321 cagccgatgc tgtatcgtct gcagcaggtt agcagtcgtc gtctgctgag caatctggtt
     1381 agcgaatttc gtcgtgaact gcctcgtgaa caggcacaag aggcaggtta tggtctggca
     1441 gcactgattg atggtctgtg gctgcgtgca gcactgagcg gtaaaccgct ggataaaacc
     1501 cgtgcaaata gcctgacccg tcattttatc acccagcatc tgccgaccga ttgataagga
     1561 tcctaattgg taacgaatca gacaattgac ggctcgaggg agtagcatag ggtttgcaga
     1621 atccctgctt cgtccatttg acaggcacat tatgcatcga tgataagctg tcaaacatga
     1681 gcagatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     1741 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     1801 gagcaaatat tttatctgag gtgcttcctc gctcactgac tcgctgcacg aggcagacct
     1861 cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     1921 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     1981 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     2041 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     2101 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     2161 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     2221 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     2281 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     2341 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
     2401 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
     2461 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     2521 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     2581 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
     2641 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg
     2701 acgctcagtg gaacgaaaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2761 accttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     2821 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     2881 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2941 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     3001 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     3061 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     3121 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     3181 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     3241 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     3301 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     3361 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     3421 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     3481 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3541 cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     3601 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtacat
     3661 cagagatttt gagacacaac caattattga aggcctccct aacggggggc ctttttttgt
     3721 ttctggtctc ccgcttaacg atcgttggct gagcgcgggt gagagggatt cgttaccaat
     3781 agacaattga ttggacgttc aatataatgc tagcag
//
