The development of ecotourism in protected areas faces the challenge of balancing conservation and ecotourism. Ecotourism suitability assessments are essential tools for managing tourism in these areas. However, current assessments often overlook biological factors, leading to adverse effects on wildlife. This study uses the Northeast China Tiger and Leopard National Park as a case study to establish a comprehensive assessment system that integrates ecotourism suitability with tiger and leopard habitat suitability, thereby linking ecotourism with wildlife conservation. The primary research methods include ecotourism suitability analysis based on the entropy weight method and habitat suitability analysis using the MaxEnt model. Based on the zoning results of ecotourism and habitat suitability, a comprehensive ecotourism suitability zoning map was produced. This map indicates that areas of very high suitability account for 45.62% of the total area, covering approximately 6152.563 km², and are primarily located on the edges of village clusters. These areas can be prioritized for developing tourism infrastructure. The comprehensive ecotourism assessment system can balance the development of ecotourism with wildlife conservation, contributing significantly to the coordinated development of economic, social, and environmental objectives.