LOCUS Exported 2755 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial NM crRNA expression: targets NM to protospacer B (ACTTTAAAAGTATTCGCCAT), p15A/chloramphenicol. ACCESSION . VERSION . KEYWORDS PM-NM!TB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2755) AUTHORS Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM TITLE Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. JOURNAL Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. PUBMED 24076762 REFERENCE 2 (bases 1 to 2755) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2755 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 239..266 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 358..444 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" promoter 665..767 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 768..1427 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(834..853) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 1437..1523 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(1813..1830) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1947..2492) /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(2027..2046) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gacaatgaaa acgttagtca tggcgcgcct tgacggctag ctcagtccta ggtacagtgc 61 tagcttaatg ctcgcacata gcagaacttt aaaagtattc gccatgttgt agctcccttt 121 ctcatttcgc agtgctacaa tccgccgcta tggtcccacg tagagcatac ggaaaaaaaa 181 gtcaaaagcc tccgaccgga ggtcggcctt acttgctagc agagtttgta gaaacgcaaa 241 aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt atggcgggcg 301 tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc ccggcggatt 361 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt 421 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca tggggagacc 481 ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg tcaggtggga 541 ccaccgcgct actgccgcca ggcaaattct gttttatcag accgcttctg cgttctgatt 601 taatctgtat caggctgaaa atcttctctc atccgccaaa acagccccgt agaaaaaggg 661 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg 721 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa 781 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat 841 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt 901 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc 961 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat 1021 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc 1081 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg 1141 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg 1201 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca 1261 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga 1321 tgccgctggc gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc 1381 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattc cggcggattt 1441 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc 1501 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagaccc 1561 cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt caggtgggac 1621 caccgcgcta ctgccgccag gcaaattctg ttttatcaga ccgcttctgc gttctgattt 1681 aatctgtatc aggctgaaaa tcttctctca tccgccaaaa cagccctcgg gacggatcgc 1741 ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat 1801 atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga 1861 aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga 1921 agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac 1981 gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg 2041 tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc 2101 attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc 2161 agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt 2221 atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc 2281 agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa 2341 ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt 2401 tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc 2461 aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaag gggtctgacg 2521 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 2581 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 2641 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcggaaccgc 2701 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc ccggc //