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N5-N10-methylene tetrahydrofolate is particularly used in different reactions: it is used by (1) thymidylate synthase for formation of 2′-deoxythymidylate, (2) N5-N10-methylene tetrahydrofolate reductase for formation of N5-methyltetrahydrofolate, and (3) N5-N10-methylene tetrahydrofolate dehydrogenase to form N5-N10-methylene tetrahydrofolate [10, 23]