Intervention in nature through the implementation of various development projects to meet the growing needs of the population is inevitable. Any intervention has both negative and positive consequences for ecosystem services; therefore, environmental impact assessment has been a focus of attention worldwide for decades. Ecosystem services valuation can serve as a complementary approach in environmental impact assessments, playing a significant role in development-related decision-making. Though, this has been ignored in the evaluation of infrastructure projects. The present study integrates environmental impact assessment approaches with the economic valuation of ecosystem services to conduct a cost-benefit analysis of the Bandar Abbas–Kerman freeway construction project. The ecosystem services considered included sediment retention, water production, habitat quality, carbon sequestration, food production, tourism, vegetation quality, forage, and oxygen production. The results (updated based on the annual inflation rate) showed that the project imposed damages (costs) of 161,842,750,785 tomans and 332,307,472,031 tomans on ecosystem services in the construction and operation phases, respectively. Similarly, no benefit observed in ecosystem services during the construction phase, while the operation phase resulted in a benefit equivalent to 366,024,608,960 tomans. Consequently, the net damage from the implementation of this project was estimated to be 128,125,613,856 tomans.