The balance between the stress rupture properties and castability of a 4th-generation Ni-based single crystal superalloy can be achieved by addition of appropriate carbon content. In this work, the effects of carbon content on the microstructure and stress rupture properties of a 4th-generation Ni-based single crystal superalloy were investigated in detail. The results showed that minor carbon addition could aggravate the dendrite segregation, while the addition of carbon had no obvious effect on the rafting and distortion of γ′ phase during stress rupture deformation. At 760 °C/810 MPa and 1100 °C/210 MPa, the stress rupture lives of the three experimental alloys initially increased and subsequently decreased with the increasing carbon content. However, under condition of 1140 °C/137 MPa, the increased carbon content resulted in the reduction of the stress rupture lives of alloy. It was discovered that minor carbon elements addition (0.045 wt%) can promote formation of interfacial dislocation networks and improve creep resistance at high temperatures. At intermediate temperature, the introduction of carbon could reduce the stacking fault (SF) energy of γ matrix, thus facilitating the formation of SFs in γ matrix. The M6Cgranular carbides precipitated at high temperature, which was considered to be beneficial to stress rupture properties.