As undergraduates' academic distress continues to rise, it is important to consider factors related to classroom anxiety. This study investigated how student anxiety varied by preferred seating location and whether the reasons for their preferred seating location were related to these anxiety levels. In seven introductory biology classes at a large public university, we assessed student anxiety levels about different classroom practices. Students also reported where they preferred to sit in a lecture class and explained why they preferred that location. We coded and created themes from these explanations without knowing the anxiety levels or student seating preferences. We compared anxiety levels among nine preferred seating locations via ANOVA: front-right, front-middle, front-left, middle-right, middle-middle, middle-left, back-right, back-middle, back-left. The back-left had higher group work anxiety. The middle-right had higher anxiety for volunteering, cold calling, and clicker use. The back-left and front-middle had higher test and clicker anxiety. Student explanations for seating preference indicated a balance between wanting to be engaged in the class yet wanting to avoid others and feel comfortable; these reasons did not directly align with anxiety differences, however. We suggest teachers consider how anxiety varies spatially in their classroom to inform their active learning implementation practices.