A microstructure-based fatigue model is employed to predict fatigue damage in 4140 steel. Fully reversed, strain control fatigue tests were conducted at various strain amplitudes and scanning electron microscopy was employed to establish structure-property relations between the microstructure and cyclic damage. Fatigue cracks were found to initiate from particles near the free surface of the specimens. In addition, fatigue striations were found to originate from these particles and grew radially outward. The fatigue model used in this study captured the microstructural effects and mechanics of nucleation and growth observed in this ferrous metal. Good correlation of the number of cycles to failure between the experimental results and the model were achieved. Based on analysis of the mechanical testing, fractography and modeling, the fatigue life of the 4140 steel is estimated to comprise mainly of small crack growth in the low cycle regime and crack incubation in the high cycle fatigue regime.