Nucleospin plasmid miniprep kit

The genes of interest were amplified from the genomic DNA of T. reesei strain PC-3-7, and a vector fragment was amplified by inverse polymerase chain reaction (PCR) using pUC118 (Takara Bio, Shiga, Japan) as the template. The amplified fragments, which have been pre-designed to add a SwaI cleavage site using primers, were ligated using an In-Fusion HD Cloning Kit (Clontech Laboratories, Mountain View, CA, USA) according to the manufacturer’s protocol. Escherichia coli DH5a was used as a cloning host, and a NucleoSpin^® Plasmid miniprep kit (Takara Bio) was used to purify the plasmid DNA. More details on the cloned gene and primers are provided in an Additional file 2: Table S2. The nanobody expression cassettes were constructed by the fragment of artificial gene synthesis (Additional file 2: Table S4) procured from the amino acid sequence of a nanobody with CBD-carrier polypeptide and KEX2 linker (Fig. 3a, SCK) shown in Additional file 2: Table S2 and the PCR fragment with the primer pair shown in Additional file 2: Table S5. The expression cassettes of secretory signal peptide (Fig. 3a, S) and CBD-carrier polypeptide (Fig. 3a, SC) addition were obtained by inverse PCR from expression cassettes of CSK patterns (pUC-V014, pUC-V017, pUC-V020) as templates.