This paper investigates the machine layout problem with a linear single-row flow path within an automated manufacturing system. Traditional machine layout approaches can produce inappropriate layout designs because they often do not consider flow path characteristics, such as flow path configuration and feasible flow path direction. This paper investigates the effects of such flow path characteristics on machine layouts. A two-phase layout procedure that combines flow line analysis with simulated annealing is proposed. The procedure can be employed using a variety of evaluation criteria (minimize total flow distance, maximize number of in-sequence movements, and so on). Different layout procedures can be obtained by varying the flow line analysis method and temperature reduction strategy employed for simulated annealing. Experiments are performed to test the performance of different layout procedures for different combinations of flow path characteristics and quantities of machines. The experimental results provide vital information on selecting appropriate flow line analysis methods and temperature reduction strategies for different layout problems. Furthermore, the results will help designers determine appropriate evaluation criteria for different layout problems.