Entrepreneurship is one of the most significant outcomes of university development and requires innovative managerial approaches such as organizational ambidexterity. Originating from industry, this approach emphasizes the simultaneous pursuit of exploiting existing capacities and exploring new knowledge for the future. This study aimed to design and validate a balanced ambidextrous university model to foster entrepreneurship. The research employed a mixed-method sequential exploratory design. In the qualitative phase, experts in higher education management were selected purposefully using the snowball technique, and 14 semi-structured interviews were conducted. In the quantitative phase, the statistical population included presidents, vice-presidents, and faculty members of comprehensive universities in Iran. Based on Cochran’s formula, 256 participants were chosen through stratified random sampling and responded to a researcher-developed questionnaire. Qualitative data were analyzed using thematic analysis, with content validity confirmed by experts and reliability ensured through re-testing. In the quantitative phase, structural equation modeling was applied to validate the model. Findings revealed six main components of exploitation: university policies, organizational structures, university–industry interaction, education–research development, budgeting, and academic culture. Exploration comprised six components: needs assessment and training, future-oriented technology and structures, financial resources, innovative culture, forward-looking management, and strategic innovation. Balancing strategies were identified in two main components: policy and evaluation, and resources and structures. The proposed ambidextrous university model, by preventing unproductive stagnation and fruitless innovation, paves the way for entrepreneurship-oriented university development. By creating effective links between education, research, and industry, it contributes to the formation of entrepreneurial ecosystems within universities.