. 2020 Sep 15;94(19):e00252-20. doi: 10.1128/JVI.00252-20

TABLE 4.

Fragments of the pCAGGS/CHIKV-E mutant plasmid

pCAGGS/CHIKV-E mutation	Primer(s) used for
pCAGGS/CHIKV-E mutation	Mutant fragment 1	Mutant fragment 2^a ^,^b ^,^d	pCAGGS/CHIKV-E vector fragment^c
YKATR/AAATA	E3-SphI-F, E2-YKR13/A-R^a ^,^b ^,^d	E2-YKR13/A-F, E2-XhoI-R	pCAGGS/CHIKV-E ΔSphI-AleI
TTTDK/AAADA	E3-SphI-F, E2-TTTK215/A-R^a ^,^b ^,^d	E2-TTTK215/A-F, E2-XhoI-R	pCAGGS/CHIKV-E ΔSphI-XhoI
HKKW/AAAA	E2-BstEII HKKW235/A-F, E2-XhoI-R^a,^d		pCAGGS/CHIKV-E ΔBstEII-XhoI
RQGK/AQGA	E3-SphI-F, E2-RK254/A-R^a ^,^b ^,^d	E2-RK254/A-F, E2-XhoI-R	pCAGGS/CHIKV-E ΔSphI-XhoI

Amplified by PCR.

Mutant fragments 1 and 2 were assembled by using an In-Fusion HD cloning plus kit.

The pCAGGS/CHIKV-E Thai#16856 plasmid was digested with the respective restriction enzyme.

The mutant fragment was digested with the respective restriction enzyme and then ligated with the respective pCAGGS/CHIKV-E vector fragment.