One defining property of stem cells is their ability to differentiate via asymmetric cell division, in which a stem cell creates a differentiated daughter cell but retains its own phenotype. Here, I describe a synthetic genetic circuit for controlling asymmetric cell division in E. coli in which a progenitor cell creates a differentiated daughter cell while retaining its original phenotype. Specifically, an inducible system was engineered that can bind and segregate plasmid DNA to a single position in the cell. Upon division, the colocalized plasmids are kept by one and only one of the daughter cells. The other daughter cell receives no plasmid DNA and is hence irreversibly differentiated from its sibling. In this way, asymmetric cell division happens though asymmetric plasmid partitioning. This system was further used to achieve physical separation of genetically distinct cells by tying motility to differentiation. Finally, an orthogonal inducible circuit was characterized that enables the simultaneous asymmetric partitioning of two plasmid species, resulting in pluripotent cells that have four distinct differentiated states. These results point the way towards engineering multicellular systems from prokaryotic hosts.