The natural system in East Kalimantan, Indonesia, is undergoing degradation, which mainly originated from the anthropogenic exploitation of nature. To reduce the harmful effects of the environmental damages, East Kalimantan students must, therefore, be examined on pro-environmental behavior (PEB) as they will be the future guardians, planners, and policymakers. This study investigated the PEB of 410 junior high school students in East Kalimantan and discussed it based on their gender and location categories (rural and urban). The investigation of PEB was conducted using an adapted version of the Two Major Environmental Values model, which includes an additional scale for measuring Appreciation. The PEB questionnaire data was analyzed quantitatively using an independent t-test to compare the means across location categories and gender. Our findings revealed that the PEB of male students (2.44) is higher than female students (2.36). Most female students favored the Utilization of nature more than male students, resulting in lower PEB. Students in rural schools were likely to behave more pro-environmentally than those in urban schools. According to the analysis of each PEB aspect, most students had Utilization preferences toward nature, especially students studying in Urban schools. These results may be valuable in designing behavioral interventions to encourage PEB, especially in East Kalimantan.