A mark for alignment and overlay, a mask having the same, and a method of using the same are provided. The mark includes a first mark pattern and a second mark pattern. The first mark pattern includes a first pattern and a second pattern, and the second mark pattern includes a third pattern and a fourth pattern. The first pattern includes a plurality of rectangular regions arranged in a first direction, and for each rectangular region, a sideline in a second direction is longer than a sideline in the first direction, wherein the first direction is perpendicular to the second direction. The second pattern is disposed on both sides of the first pattern in the second direction and includes a plurality of rectangular regions arranged in the second direction, and for each rectangular region, the sideline in the first direction is longer than a sideline in the second direction. The third pattern includes two rectangular regions disposed on both sides of the first pattern in the first direction, and the fourth pattern includes two rectangular regions disposed on both sides of the second pattern in the second direction.