A DNA sequence outside the pUB110 minimal replicon is required for normal replication in Bacillus subtilis

The origin of lagging strand synthesis in pUB110, oriL, has been localized within 140 bases outside the pUB110 minimal replicon. The oriL DNA sequence is a cis-acting and orientation dependent determinant required for normal plasmid replication. Rearrangements affecting oriL cause plasmid instability, lead to the accumulation of replication intermediates and result in a marked reduction of the plasmid copy number in some recombination deficient mutant strains. In addition, deletion of oriL triggers a dnaB-dependent mode of replication. Insertion of the functionally asymmetric oriL region in the proper orientation into pC194 reduces the accumulation of single-stranded DNA during the replication of this plasmid.