F plasmid is partitioned with fidelity to daughter cells during cell division cycle owing to two trans-acting genes, sopA and sopB, and a cis-acting site, sopC. We visualized the subcellular distribution of mini-F-plasmid molecules by fluorescence in situ hybridization. Mini-F-plasmid molecules having the sopABC segment were localized at midcell in newborn cells. Replicated plasmid molecules migrated to cell positions 1/4 and 3/4 without coupling with cell elongation and were tethered to these positions until completion of cell division. In contrast, molecules of a mini F plasmid lacking the sopABC segment were distributed randomly in spaces not occupied by nucleoids. The sopABC system caused replicated plasmid molecules to be positioned and tethered at the cell quarter sites.