LOCUS Exported 4098 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PcaUAM + P3B5B-YFP Reporter. DHBA sensor for use in non-Marionette strain.. ACCESSION . VERSION . KEYWORDS pAJM.690 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4098 /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(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 /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 //