Using fluorescence spectroscopy we directly measure entropy production of a single two-level system realized experimentally as an optically driven defect center in diamond. We exploit a recent suggestion to define entropy on the level of a single stochastic trajectory [Seifert, Phys. Rev. Lett. 95, 040602 (2005)10.1103/PhysRevLett.95.040602]. Entropy production can then be split into one of the system itself and one of the surrounding medium. We demonstrate that the total entropy production obeys various exact relations for finite time trajectories.