Construction of the integrative vector pUE10

A plasmid derived from pCB1004 [38] (pUE07) containing a chloramphenicol resistance gene and a hygromicin B resistance gene was used as the base for the construction of pUE10 [39]. pUE07 contains the E. coli f1 and colE1 origins, as well as a multiple cloning site and the trpC terminator of Aspergillus nidulans.

The trpC terminator sequence in pUE07 was replaced by a sequence containing the blu17 terminator and a region of 804 bp downstream of it (to direct integration by homologous recombination), by digestion with XhoI and KpnI. The promoter of the pki1 gene from T. reesei was inserted in the SacI and SacII sites (S1 Fig). Two other versions of this vector with distinct inducible promoters (pbr1 from T. atroviride and qa2 from Neurospora crassa) were also constructed (data not shown).