Nowadays, high-throughput screening (HTS) systems are widely used in pharmaceutical industries and laboratories for the discovery of new drugs and biomedical substances. It is important to efficiently schedule them so as to reduce the cost. In the operation of an HTS system, a microplate may visit some resources more than once and complex time window constraints are imposed on some activities and activity sequences. Moreover, with the consistency requirement, a one-microplate cyclic schedule is necessary. Thus, its scheduling problem is very challenging. This article studies the scheduling problem of an HTS system for an enzymatic assay, a typical application of HTSs, from the perspective of control theory. The system is modeled by resource-oriented Petri nets (ROPNs). With the model, necessary and sufficient conditions under which a feasible cyclic schedule exists are established. Then, we determine how many microplates should be in the system for concurrent processing and the transition firing sequence to obtain the activity sequence for an optimal and feasible schedule. In this way, a feasible and optimal cyclic schedule can be found by very simple computations, which shows that polynomial algorithms for an optimal schedule exist, while the existing studies apply mathematical programming with exponential computation complexity. Also, its efficient implementation is given.