Recently, the single-shaft series-parallel powertrain of Plug-in Hybrid Electric Bus (PHEB) has become one of the most popular powertrains due to its alterable operating modes, excellent fuel economy and strong adaptability for driving cycles. Nevertheless, for configuring the PHEB with single-shaft series-parallel powertrain in the development stage, it still faces greater challenge than other configurations when choosing and matching the main component parameters. Motivated by this issue, a comprehensive multi-objectives optimization strategy based on Genetic Algorithm (GA) is developed for the PHEB with the typical powertrain. First, considering repeatability and regularity of bus route, the methods of off-line data processing and mathematical statistics are adopted, to obtain a representative driving cycle, which could well reflect the general characteristic of the real-world bus route. Then, the economical optimization objective is defined, which is consist of manufacturing costs of the key components and energy consumption, and combined with the dynamical optimization objective, a multi-objective optimization function is put forward. Meanwhile, GA algorithm is used to optimize the parameters, for the optimal components combination of the novel series-parallel powertrain. Finally, a comparison with the prototype is carried out to verify the performance of the optimized powertrain along driving cycles. Simulation results indicate that the parameters of powertrain components obtained by the proposed comprehensive multi-objectives optimization strategy might get better fuel economy, meanwhile ensure the dynamic performance of PHEB. In contrast to the original, the costs declined by 18%. Hence, the strategy would provide a theoretical guidance on parameter selection for PHEB manufacturers.