An accurate large-scale positioning system is a high-precision distributed large-scale dimensional metrology system that is widely used in smart manufacturing and assembly applications. The system comprises multiple transmitters with locations and orientations that are typically calibrated based on auxiliary measurement equipment. However, the calibration method is complicated and time consuming. We propose a rapid and flexible calibration method based on the highly precise three-dimensional coordinate control network established by a calibration bar with an accuracy of   ±  0.005  mm, including both estimation and optimization algorithms. Exploratory experiments were performed to validate the feasibility of the proposed calibration method. The results demonstrate that the proposed calibration method enables the repeated measurement errors to be maintained at <0.05  mm in X and Y and <0.04  mm in Z. The standard deviation after calibration was 0.18 mm in comparison with the laser tracker.