Regulatory elements coordinating initiation of chromosome replication to the Escherichia coli cell cycle doi-10-17044-scilifelab-22139918-0 https://doi.org/10.17044/SCILIFELAB.22139918 Swedish National Data Service Svensk nationell datatjänst Landing page Regulatory elements coordinating initiation of chromosome replication to the Escherichia coli cell cycle doi-10-17044-scilifelab-22139918-0 https://doi.org/10.17044/SCILIFELAB.22139918 Broström, Oscar Fange, David Swedish National Data Service Svensk nationell datatjänst Landing page Bacteria replicate the genome on average once per generation, irrespective of growth condition-dependent changes in generation time and cell size. The strategy appears to be to initiate replication in a narrow chromosome-to-size ratio interval for each growth condition. To investigate how Escherichia coli accurately compares its size to the number of chromosomes, we perturbed some of the mechanisms proposed to achieve this precision and monitored how replication initiation was affected with time-lapse single-cell fluorescence microscopy. We found that the regulatory inactivation of DnaA-ATP (RIDA) mechanism, which converts the replication initiation molecule DnaA to its inactive form in proportion to the number of chromosomes, was the most important factor and that other previously described mechanisms have a less critical role.  This dataset contains the raw data, analysis and code needed to generate the figures presented in the paper. The raw data consists of microscopy images, growth rate measurements, RT-qPCR, whole-genome sequencing and Rif-runout data. The analysis and code part consists of code and output from the analysis and post-processing. Access to data through an external actor. Åtkomst till data via extern aktör.