Significance Our innate circadian clocks control myriad aspects of behavior and physiology. Disruption of our clocks by shift work, jet lag, or inherited mutation leads to metabolic dysregulation and contributes to diseases, including diabetes and cancer. A central step in clock control is phosphorylation of the PERIOD 2 (PER2) protein. Here we conclusively identify the elusive PER2 priming kinase and find it to be the well-known circadian kinase, casein kinase 1 (CK1). Surprisingly, different forms of CK1 have differing abilities to phosphorylate the PER2 priming site, adding to the complexity of circadian regulation. These insights into the phosphoregulation of PER2 will be of broad interest to circadian biologists, computational modelers, and those seeking to pharmacologically manipulate the circadian clock.