LOCUS Exported 3863 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION TetR + PTet*-YFP Reporter. aTc sensor for use in non-Marionette strain.. ACCESSION . VERSION . KEYWORDS pAJM.011 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3863) 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 3863) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3863 /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" CDS 967..1590 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind complement(1040..1059) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(1780..1799) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" rep_origin complement(2164..2709) /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(2244..2263) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2798..3613) /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 3521..3540 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3605..3624 /label=pENTR-R /note="pENTR vectors, reverse primer" protein_bind 3808..3826 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3833..3851 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" 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 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg 961 gtgaatatgt ccagattaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat 1021 gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag 1081 cctacattgt attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag 1141 atgttagata ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt 1201 ttacgtaata acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa 1261 gtacatttag gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc 1321 tttttatgcc aacaaggttt ttcactagag aatgcattat atgcactcag cgctgtgggg 1381 cattttactt taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg 1441 gaaacaccta ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat 1501 caccaaggtg cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa 1561 aaacaactta aatgtgaaag tgggtcctga taaggatcct aattggtaac gaatcagaat 1621 tgacggctcg agggagtagc atagggtttg cagaatccct gcttcgtcca tttgacaggc 1681 acattatgca tcgatgataa gctgtcaaac atgagcagat cctctacgcc ggacgcatcg 1741 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg 1801 atggggaaga tcgggctcgc cacttcgggc tcatgagcaa atattttatc tgaggtgctt 1861 cctcgctcac tgactcgctg cacgaggcag acctcagcgc tagcggagtg tatactggct 1921 tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg 1981 ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg 2041 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag 2101 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc 2161 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg 2221 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt 2281 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt 2341 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta 2401 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt 2461 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag 2521 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac 2581 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa 2641 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa 2701 cgatctcaag aagatcatct tattaagggg tctgacgctc agtggaacga aaaatcaatc 2761 taaagtatat atgagtaaac ttggtctgac agttacctta gaaaaactca tcgagcatca 2821 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt 2881 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc 2941 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa 3001 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa 3061 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat 3121 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc 3181 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg 3241 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg 3301 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct 3361 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa 3421 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc 3481 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc 3541 catataaatc agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt 3601 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc 3661 atgatgatat atttttatct tgtgcaatgt acatcagaga ttttgagaca caaccaatta 3721 ttgaaggcct ccctaacggg gggccttttt ttgtttctgg tctcccgctt aacgatcgtt 3781 ggctgttttc agcaggacgc actgacctcc ctatcagtga tagagattga catccctatc 3841 agtgatagag atactgagca cag //