In this paper, the driving data of heavy semi-trailer tractors on six round routes were collected by the vehicle driving data platform of SINOTRUK, and the statistical characteristics of the database were calculated. The Markov chain method was used to construct driving cycles in the comprehensive routes of heavy semi-trailer tractors, and the driving cycles were optimized with the objective of the deviation of the statistical characteristic indexes of the driving cycles and the database by the genetic algorithm, which improved the accuracy of the results. Fuel consumption simulation was performed to verify the design results, and the deviation between the calculated and actual values was within the satisfactory range, and the design cycles could represent the fuel consumption of the vehicle traveling in a specific area. Comparison of the design results with the standard cycles CHTC-TT and C-WTVC shows that the design results are more representative of the driving characteristics of heavy tractors on common routes. The comprehensive driving cycle design method applied in this paper can help to construct high-precision driving cycles that reflect the typical driving characteristics of semi-trailer tractors in actual areas, and provide a more accurate benchmark for the development of vehicle economy and other work.