Abstract The ability to perceive spatial environments and locate oneself during navigation is crucial for the survival of animals. Mounting evidence suggests a role of the medial prefrontal cortex (mPFC) in spatially related behaviors. However, the properties of mPFC spatial encoding and how it is influenced by animal behavior are poorly defined. Here, we train the mice to perform 3 tasks differing in working memory and reward-seeking: a delayed non-match to place (DNMTP) task, a passive alternation (PA) task, and a free-running task. Single-unit recording in the mPFC shows that although individual mPFC neurons exhibit spatially selective firing, they do not reliably represent the animal location. The population activity of mPFC neurons predicts the animal location. Notably, the population coding of animal locations by the mPFC is modulated by animal behavior in that the coding accuracy is higher in tasks involved in working memory and reward-seeking. This study reveals an approach whereby the mPFC encodes spatial positions and the behavioral variables affecting it.