Temporarily storing sequences of events (sequence memory, SM), a fundamental ability in cognition, involves item-by-item serial reactivations during retention, yet the causal evidence is lacking. Here we develop a "behavioral temporal interference" approach to manipulate the item-specific reactivation and replay profile in the delay period when human subjects hold a list of items in working memory (WM). Eight experiments on more than two hundred subjects demonstrate that the manipulation

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... fully alters SM behavior. When items in the list are synchronized with each other and thus could not elicit time-dissociated responses, the serial position effect (SPE), a typical SM behavior, is disrupted. Reversing the temporal order of reactivations also alters SPE, supporting that the replay order is crucial as well. Our findings support the causal role of temporally ordered reactivation in mediating SM, and also provide a promising, efficient approach to manipulate the temporal structure of multiple items held in WM.