Automated vehicles (AVs) will likely influence various aspects of the existing transportation system in the next few decades. Existing literature documents the operational benefits of AVs at different penetration rates. However, estimating the effect of the mix of different levels of AVs on the operational performance of traffic flow has not been explored in the past. This study employs a microsimulation approach to model and estimate the potential effect of varying levels of AVs on the operational performance of freeways. The proposed approach involves a two-step process for calibrating the simulation model. In the first step, the model is calibrated from an operational perspective. In the second step, the calibration is performed from a safety perspective. Thirteen scenarios were generated considering varying penetration rates of different levels of AVs. Congestion index (CI), buffer time (BT), travel time uncertainty (TTU), and delay were computed for each scenario and segment type. The estimations indicate that increasing the penetration of level 3 to level 5 AVs significantly reduces CI, BT, TTU, and delay. In contrast, the effect of level 1 and 2 AVs on freeway operational performance is negligible. Notably, the operational benefits are more pronounced with higher penetration of level 3 and higher AVs. Additionally, the reduction in delay is greater on interchange segments compared with basic freeway segments. The study findings are valuable insights concerning the operational performance of freeways under varying penetration of different levels of AVs, assisting practitioners in formulating AVs-inclusive policies.