Table 1.

Primers used in the study (restriction sites represented in upper case)

Construct name (intermediate construct)	Vector	Forward primer (5’–3’)	Reverse Primer (5’–3’)	Linearized vector	Primary PCR templatea	Secondary PCR templatea
BglA-v (BglA)	pET21a	CTTTAAGAAGGAGATATACATATGGCTAGCcaccatcaccatcaccacaagataactttcccaaaaga	ccatgtaggaacgagctttgtgccACTAGTCTGAGGAGTTGTTACAGTTGTgaaaccgttgtttttgattacttc		pET28a BglA [32]	pET21a
BglA-v	pET21a	ttacatACTAGTtctcataaatttatctatgg	ttactaGAGCTCctattgttcttcaactggga	pET9d Xyn-docB [48]		–
BglA*-v (BglA*)	pET21a	CTTTAAGAAGGAGATATACATATGGCTAGCcaccatcaccatcaccacaagataactttcccaaaaga	ccatgtaggaacgagctttgtgccACTAGTCTGAGGAGTTGTTACAGTTGTgaaaccgttgtttttgattacttc		pET28a BglA*	pET21a
BglA*-f	pET21a	CTTTAAGAAGGAGATATACATATGGCTAGCcaccatcaccatcaccacaagataactttcccaaaaga	ccatgtaggaacgagctttgtgccACTAGTCTGAGGAGTTGTTACAGTTGTgaaaccgttgtttttgattacttc		pET28a BglA*	pET21a Xyn10A-f [42]
BglA*-v	pET21a	ttacatACTAGTtctcataaatttatctatgg	ttactaGAGCTCctattgttcttcaactggga	pET9d Xyn-docB [48]		–
8A-g (8A)	pET28a	ttacgtCCATGGgtgtgccttttaacacaaaata	ttacgaGGTACCaatgaaggtgtcggattcga	pET28a Cel8A [29]		–
8A-g	pET28a	tactagGGTACCagaagaagcaaacaagggag	ttactaCTCGAGcttacccagtaagccattct	pET28a-g-5A [6]		–
8A*-g (8A*)	pET28a	ttacgtCCATGGgtgtgccttttaacacaaaata	ttacgaGGTACCaatgaaggtgtcggattcga	pET28a 8AQM [24]		–
8A*-g	pET28a	tactagGGTACCagaagaagcaaacaagggag	ttactaCTCGAGcttacccagtaagccattct	pET28a-g-5A [6]		–
8A*-b	pET28a	ttacgtCCATGGgtgtgccttttaacacaaaata	ttacgaGGTACCaatgaaggtgtcggattcga	pET28a 8A-b [29]
Scaf·GT	pET9d	GTTTAACTTTAAGAAGGAGATATAccatggtgcttcctccgaaaactacc	ggtgtgtttgtcggtgtgtttgtcGGTACCgcttcttcctgagagacaatc		pET28a CBM-Coh2375 [34]	pET28a Scaf·AT [49]
Scaf·GTV	pET9d	gttctttgacggtggagtaaatgttggagatacaacagtacctacaacacctacaacacctGCAGGGCAATTACAAATTGA	CCTTTCGGGCTTTGTTAGCAGCCggatcctAATTTGAGCCAACCAATATAG		pET28a CBM-CohC1 [48]	pET9d Scaf·GT

^aIndicates restriction-free cloning