LOCUS Exported 3816 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION BetIAM + PBetI-YFP Reporter. Cho sensor for use in non-Marionette strain.. ACCESSION . VERSION . KEYWORDS pAJM.683 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3816 /organism="synthetic DNA construct" /mol_type="other DNA" misc_RNA 1..50 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" CDS 100..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 /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 //