The errors of the overlay printing registration in roll-to-roll (R2R) gravure are generated from many sources such as uneven web tension, thermal expansion and contraction of plastic web, uneven impression roll pressure, slipping the web, uneven circumference of the roll, unmatched speed between web transfer, printing roll, and so forth. Among those things, the most influential factors to exert on overlay printing registration in R2R gravure should be defined and analyzed to provide servomechanism to control at least ±20 µm of registration accuracy of R2R gravure with more than 4 m/min of web transfer speed. In this paper, we would like to present the general way of control system of R2R gravure to maintain the overlay printing registration of ±20 µm under various web transfer speeds, roll pressure and uneven web tension