In order to improve the efficiency of assembly line and optimize the layout, this paper presents a collaborative optimization model for a two-sided U-type assembly line and a novel design with p-l partition layout is proposed to minimize number of workstations without increasing the length of the assembly line. Considering the task orientation and time sequencing in cross-workstation, the mathematical model of two-sided U-type assembly line balancing problem is derived. A multi-level priority rule heuristic algorithm is developed to drive the optimization process. The multi-level priority rule heuristic algorithm, modified particle swarm optimization algorithm, and the bi-objective integer programming method are applied to 20 classic examples, respectively. The calculation results suggest that the optimal results of the proposed method account for 90%, which verifies the rationality of the collaborative optimization model and algorithm, and provides a useful reference for the modeling and solution of the two-sided U-type assembly line balancing problems.