The Evolutionary Attitudes and Literacy Survey (EALS) is a multidimensional scale consisting of 16 lower- and 6 higher-order constructs developed to measure the wide array of factors that influence both an individual’s endorsement of and objection to evolutionary theory. Past research has demonstrated the validity and utility of the EALS (Hawley et al., Evol Educ Outreach 4:117–132, 2011); however, the 104-item long-form scale may be excessive for researchers and educators. The present study sought to reduce the number of items in the EALS while maintaining the validity and structure of the long form. For the present study, and following best practices for short-form construction, we surveyed a new sample of several hundred undergraduates from multiple majors and reduced the long form by 40% while maintaining the scale structure and validity. A multiple-group confirmatory factor analysis supported strong factorial invariance across samples, and therefore verified structure and pattern between the six higher-order constructs of the long-form EALS and the EALS short form (EALS-SF). Regression analysis further demonstrated the short form’s validity (i.e., demographics and openness to experience) and replicated previous findings. In the end, the EALS-SF may be a versatile tool that may be used whole or in part for a variety of research areas, including curricular effectiveness of courses on evolution and/or biology.