In this study, a template polymer with anionic microblock structure was successfully synthesized through ultrasonic initiated template copolymerization (USTP) by using sodium allylsulfonate (SAS) and acrylamide (AM) as monomers, poly diallyl dimethyl ammonium chloride (polyDADMAC) as template, and 2,2′-azobis [2-(2-imidazolin-2-yl) propane] dihydrochloride (VA-044) as initiator. The anionic polyacrylamide (APAM-T) flocculant was characterized by Fourier transform infrared spectroscopy (FTIR), 1H nuclear magnetic resonance spectroscopy (1H NMR), and scanning electron microscopy (SEM). Furthermore, the X-ray photoelectron spectroscopy (XPS) was conducted to measure the elemental composition. The results of XPS and 1H NMR spectroscopy analysis indicated the existence of anionic microblock structure in APAM-T. Besides, the copolymerization was revealed to follow the self-assembly mechanism by analyzing the association constant (KM). The flocculation performance was evaluated by turbidity and zeta potential in the flocculation tests. The results indicated that the APAM-T with microblock structure enhanced the charge neutralization and bridging adsorption capacity, thus improving the flocculation performance. In the floc particle size distribution and fragmentation and reflocculation experiments, the floc corresponding to APAM-T had the strongest regeneration ability after fragmentation, which further confirmed the criticality of the microblock structure in enhancing the flocculation performance. Compared with existing studies, this study not only demonstrated a novel synthesis method, but also found that the microblock structure could significantly improve its treatment efficiency and effectiveness, providing a new theoretical basis and practical application path for wastewater treatment technology.