We describe here the development and validation of the Academic Career Readiness Assessment (ACRA) rubric, an instrument that was designed to provide more equity in mentoring, transparency in hiring, and accountability in training of aspiring faculty in the life sciences. We report here the results of interviews with faculty at 20 U.S. institutions which resulted in the identification of 14 qualifications and levels of achievement required for obtaining a faculty position at three groups of institutions: research-intensive (R), teaching-only (T), and research and teaching-focused (RT). T institutions hire candidates on teaching experience and pedagogical practices, and on their ability to serve diverse student populations. RT institutions hire faculty on both research and teaching-related qualifications, as well as on the ability to support students in the laboratory. R institutions hire candidates mainly on their research achievements and potential, which may limit the diversification of the life science academic pathway.