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In Escherichia coli, the DnaB helicase forms the basis for the assembly of the DNA ****tion complex. The stability of DnaB at the ****tion fork is likely important for successful ****tion initiation and progression. Single-molecule experiments have significantly changed the classical model of highly stable ****tion machines by showing that components exchange with free molecules from the environment. However, due to technical limitations, accurate assessments of DnaB stability in the context of ****tion are lacking. Using i