Genomes are replicated by large protein complexes called replisomes. In bacterial DNA replication, two replisomes replicate the DNA starting from the same origin site and proceeding in opposite directions. Understanding their movement in vivo has been challenging. We used quantitative genome sequencing to characterize the dynamics of bacterial replisomes at 5 different temperatures (17, 22, 27, 32 and 37 °C) in exponential growth (3 replicates) or in stationary phase (one experiment at 17, 27 and 37 °C). The data deposited here give the coordinates of the sequence reads (deposited under the BioProject PRJNA772106) covering the Escherichia coli str. K-12 substr. MG1655 complete genome (accession number U00096.3). The file archive contains data files for each sample, at nucleotide resolution and binned in intervals of 10,000 base pairs. It also contains a C program to perform the binning and a README summarising how the alignment was done. Please note that once uncompressed, the data will take 5 Gb of disks space in total.