The electrification of bus-based public transportation contributes to the goal of reducing the adverse environmental impacts caused by urban transportation. However, the penetration of electric vehicles has been slow due to their lower vehicle range and total costs in comparison to vehicles driven by internal combustion engines. By improving the powertrain efficiency, the total costs can be reduced for the same vehicle range. Therefore, this paper proposes a holistic design exploration approach to investigate and identify the optimal powertrain concept for electric city buses based on the component costs and energy consumption costs. The load profiles of speed, slope, and passenger occupancy profiles are derived for a selected bus route in Singapore, which is used in a powertrain design exploration for a 30-passenger vehicle. Six different powertrain architectures are analyzed, together with single and multi-speed gearbox configurations, to identify the optimal powertrain architecture and the resulting component sizes. The powertrain configurations are further analyzed in terms of their influence on the vehicle characteristics and total costs. Multi-motor configurations were found to have better vehicle characteristics and lower total costs in comparison to single rear motor configurations. Concepts with motors on the front and a rear axle could shift the load points to a higher efficiency region, resulting in lower energy consumption and energy costs. The optimal powertrain concept was a fixed-speed two-motor configuration, with a booster motor on the front axle and a motor on the rear axle.