As a novel antimicrobial material, quaternary phosphonium salts (QPSs) have been drawing close attention because of their excellent antimicrobial capacity with high activity and low bacterial survivability. Polymeric QPSs (PQPSs) also exhibit selectivity and long-term stability, however the polymerization of QPSs is severely challenged by low controllability and narrow selectivity of cation type. In this study, high-conversion RAFT polymerization is employed to prepare innovative phosphonium-functionalized diblock copolymers (PFDCs) with desired molecular weights and particle sizes. The excellent antibacterial activity of the PFDCs achieves lowest MIC values of 40 and 60 μg mL⁻¹ (i.e., 1.4 and 2.2 μmol L⁻¹) against E. coli and S. aureus, respectively. Mixing with an ink, dye, and latex coating does not weaken the antibacterial activity of the PFDCs, which inhibited 99.9% E. coli, showing broad applicability in different media. The effects of the cation type, synthesis medium, crosslinking content, and particle size on the morphology and antibacterial activity are studied. In summary, the RAFT polymerization of QPSs through the versatile design of ionic liquid monomers and the polymerization-induced self-assembly (PISA) method for constructing nano-assemblies with various micromorphology and particle size provides an exceedingly efficient way to build up multifunctional and multi-morphological polymeric nano-objects that open up vast possibilities in the fields of antibiotics, drug delivery, templated synthesis, and catalysis.