License plate recognition (LPR) data are emerging data sources that provide rich information in estimating the traffic conditions of urban arterials. While large-scale LPR system is not common in US, last few years have seen rapid developments and implementations in many other parts of world (e.g. China, Thailand and Middle East). Due to privacy issues, LPR data are seldom available to research communities. However, when available, this data source can be valuable in estimating real-time operational metrics in transportation systems. This paper proposes a lane-based real-time queue length estimation model using the license plate recognition (LPR) data. In the model, an interpolation method based on Gaussian process is developed to reconstruct the equivalent cumulative arrival–departure curve for each lane. The missing information for unrecognized or unmatched vehicles is obtained from the reconstructed arrival curve. With the complete arrival and departure information, a car-following based simulation scheme is applied to estimate the real-time queue length for each lane. The proposed model is validated using ground truth information of the maximum queue lengths from the city of Langfang in China. The results show that the model can capture the variations in queue lengths in the ground truth data, and the maximum queue length for each signal cycle can be estimated with a reasonable accuracy. The estimated queue length information using the proposed model can serve as a useful performance metric for various real-time traffic control applications.