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Transcription by RNA polymerase (RNAP) is interrupted by pauses that play diverse regulatory roles. Although individual pauses have been studied in vitro, the determinants of pauses in vivo and their distribution throughout the bacterial genome remain unknown. Using nascent transcript sequencing, we identified a 16-nucleotide consensus pause sequence in(More)
Supplemental Experimental Procedures Strains All experiments were performed with E. coli K12 strain MG1655 or an isogenic strain encoding three copies of the haemagglutinin (HA) epitope tag at the 5énd of nusG (MG1655 HA 3 ::nusG; RL1664). This strain was constructed by λ Red-mediated recombination without selection using a suicide plasmid encoding HA 3(More)
Essential gene functions underpin the core reactions required for cell viability, but their contributions and relationships are poorly studied in vivo. Using CRISPR interference, we created knockdowns of every essential gene in Bacillus subtilis and probed their phenotypes. Our high-confidence essential gene network, established using chemical genomics,(More)
Recombinant DNA technology has revolutionized biomedical research with continual innovations advancing the speed and throughput of molecular biology. Nearly all these tools, however, are reliant on ​ Escherichia coli​ as a host organism, and its lengthy growth rate increasingly dominates experimental time. Here we report the development of ​ Vibrio(More)
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