An object is to suppress reading error even in the case where writing and erasing are repeatedly performed. Further, another object is to reduce writing voltage and erasing voltage while increase in the area of a memory transistor is suppressed. A floating gate and a control gate are provided with an insulating film interposed therebetween over a first semiconductor layer for writing operation and erasing operation and a second semiconductor layer for reading operation which are provided over a substrate; injection and release of electrons to and from the floating gate are performed using the first semiconductor layer; and reading is performed using the second semiconductor layer.