Efficient sequential repetitive gene deletions in Neurospora crassa employing a self-excising β-recombinase/six cassette

► Demonstrated the functionality of a bacterial recombination system in N. crassa. ► The mus-51 deletion strain was successfully generated using this system. ► The marker was successfully recycled. ► The second gene cre-1 was subsequently deleted using the knock-in cassette using the same marker. ► Only one transformation is needed for one deletion and marker recycling.