. 2020 Feb 26;13:31. doi: 10.1186/s13068-020-1669-3

Table 4.

PCR primers and templates used to prepare plasmid constructs

Primer pair	Sequence (5′ to 3′)	Template
PyrG disruption cassette
pyrG-up	tcgagctcggtacccccagaggtgactttatccaagattcc	AOK27L
pyrG-up	cccgggcaattgccgcgaaaaattaaattgaatc	AOK27L
pyrG-down	cggcaattgcccggggtagtggtggatacgtactccttttatg	AOK27L
pyrG-down	ctctagaggatccccttcaggtcacgttctaagcttatcagc	AOK27L
LigD deletion cassette
ligD-up	tcgagctcggtacccggttactgctctcccttgatgatg	AOK27L
ligD-up	taggtagtgaacctatttcgagagcag	AOK27L
ligD-down	taggttcactacctagcggccgcacaggcaccttgcatcatcatc	AOK27L
ligD-down	ctctagaggatccccggaccgacgattcgttgaagag	AOK27L
ligD-up2	acaggtatcgaattcgtccttgtgacgacgagctcg	AOK27L
ligD-up2	ggtgcctgtgcggccgctaggtagtgaacctatttcgagagcag	AOK27L
An_pyrG set	gaattcgatacctgtcgaaagaaatggaag	FGSC-A4
An_pyrG set	cactacctagcggcctcagtgcttgtctaccagattagggag	FGSC-A4
Enzyme production cassettes
pyrG-up2	actagtcatatggatccagaggtgactttatccaagattcc	AOK27L
pyrG-up2	gtagacaagcactgacaattgccgcgaaaaattaaattgaatctatgg	AOK27L
An_pyrG-T set	acccggggatccgatgaattcgatacctgtcgaaagaaatggaag	FGSC-A4
An_pyrG-T set	Tcagtgcttgtctaccagattaggg	FGSC-A4
pyrG-down-T	gacagtcgtcgttgggtagtggtggatacgtactccttttatgg	AOK27L
pyrG-down-T	tcgagctcggtacccttcaggtcacgttctaagcttatcagc	AOK27L
P-enoA142	ccaacgacgactgtctcattactagtc	OZ
P-enoA142	attcatcggatcccgggtttgcgagtggtttggtaaaaggtag	OZ
T-agdA	gaagcgtaacaggatagcctagacc	AOK27L
T-agdA	aattcatcggatcccagtaacccattcccggttctctag	AOK27L
cbh	accactcgcaaacccatgtctgccttgaactctttcaatatgtacaag	H1
cbh	atcctgttacgcttcctacaaacattgagagtagtaagggttcacg	H1
bgl	accactcgcaaacccatgtactccgcctttctgctc	H1
bgl	atcctgttacgcttctcactggaggcactgggag	H1
ex	accactcgcaaacccatggtccgccctaccatcc	SG
ex	atcctgttacgcttctcactgctggagatcctggac	SG

Templates are genomic DNAs purified from the indicated strains. AOK27L: A. oryzae strain AOK27L, FGSC-A4: Aspergillus nidulans strain FGSC-A4, OZ: A. oryzae strain OZ, H1: T. cellulolyticus strain H1, SG: T. aurantiacus strain SG