A display substrate and a method of manufacturing a display substrate are disclosed. The display substrate includes an active pattern, a first gate electrode and a second gate electrode. The active pattern is disposed on a base substrate. The first gate electrode overlaps the active pattern. The first gate electrode is spaced apart from the active pattern by a first distance. The second gate electrode overlaps the active pattern. The second gate electrode is spaced apart from the active pattern by a second distance which is larger than the first distance.