TABLE 1.

Oligonucleotides used for plasmid construction

Mutation	Polarity	Oligonucleotide sequence (5′→3′)
Site-directed mutagenesis
pET22→pET22 LS−	−	CTGCTGTGTAGACTCCATGGTATATCTCCTTCTTAAAG
NSP3A→NSPA4–178	−	CGATTCAAATCGTTTCTCGAGTTGTTCATATCT
NSP3A→NSP3A163–313	−	CAACTTCCATGGTTTCTTCAAC
NSP3A→NSP3A206–313	−	GTTTTGGAGAGAGTCCATGGTCTCGAGTACCTTTTTTGC
NSP3A→NSP3A238–LESM (NSP3A4–237, NSP3A242–313)	−	GACTGAAGACACCATGGACTCGAGTTTATTTTGTAG
NSP3A→NSP3A10–313	−	AATCGAACTAGCCATGGTATATCTCCTTCTTAA
NSPA4–178→NSPA4–149	−	GTGGTGCTCGAGACGTTCAATCTTATCTCT
NSPA4–178→NSPA4–130	−	GTGGTGCTCGAGTGGAACTCCTTCACATT
NSPA4–178→NSPA4–178 MV54	−	AACACCTGAATCGTCGACTACGTAATCAAA
NSPA4–178→NSPA4–178 KL106	−	AGATGATAGCATAAGCTTAAGTTTGTTCAC
NSPA4–178→NSPA4–178 GS35	−	TTCATTATAATCATAGGATCCGCCCATAAGTTC
NSPA4–178→NSPA4–178 QE172	−	TTCAAATCGTTTTTCGAGCTCTTCATATCT
NSP3C→NSPC1–170	−	ATCAATCATACACTCGAGTTCTTCTATCTT
NSPC1–170→NSPC1–140	−	GTGGTGCTCGAGTACTGCATTTTCGTTTAA
PCR
NSPA163–313→NSPA163–290	+	GTTGAAGAAGCCATGGAAGTTGAC
	−	TAGCCGCTCGAGTATAATATTTCT
Insertion of sequences
pGBT9→pGBT9-NS	+	AATTCGCCATGGATCCGTCGACTGACTGCA
	−	GTCAGTCGACGGATCCATGGCG