Transportation 5.0 is a paradigm that brings the human-centered, sustainable, and smart systems approach of Industry 5.0 to the transportation sector. As transportation systems evolve towards Transportation 5.0, conceptual frameworks are required to assess and evaluate their readiness and development. This study addresses this research gap by examining Industry 4.0, Industry 5.0, and Transportation 5.0 from a broad perspective and by reviewing existing studies on Transportation 5.0. Based on the findings, a new and updated framework is proposed. The proposed framework is designed to assess the extent to which transportation systems align with the goals of Transportation 5.0, such as sustainability, digitalization, and human-centered mobility. Therefore, the various dimensions and criteria of the proposed framework are prioritized and ranked using an Analytical Hierarchical Process (AHP) method, based on expert decision-makers’ evaluations. Thus, our study supports decision-makers in their investment and policy decisions during the adoption and implementation of Transportation 5.0 paradigm and serves as a foundation for future research.