Table 1.

Primers used in this study.

Primers	Sequences (5′-3′)	Usage
Fitness-U	AGGACCGGTGAAGAAACCTGTCG	For 3tness assay
Fitness-L	AAATAGGAAGCGGGAAAAGCC
Frequency-U	CTGACCCCGCCTACGGGAAAGTG	For mutation frequency assay
Frequency-L	AGTAGTAAGTGGCAGTACGGAGG
3D-D5N-U	CGAGGGATTGATAGTTAACACCAGAGATGTTGAG	Production of the D5N mutant
3D-D5N-L	CTCAACATCTCTGGTGTTAACTATCAATCCCTCG
3D-A38V-U	GAATTTGGGCCTGCCGTCTTGTCCAACCAG	Production of the A38V mutant
3D-A38V-L	GTCCTGGTTGGACAAGACGGCAGGCCCAAATTC
3D-M194I-U	CATTCTTTACACCAGGATTATGATTGGTAGATTTT	Production of the M194I mutant
3D-M194I-L	GCAAAATCTACCAATCATAATCCTGGTGTAAAGAA
3D-M296V-U	CACTGTTGAAGGCGGGGTGCCGTCTGGTTGTT	Production of the M296V mutant
3D-M296V-L	GAACAACCAGACGGCACCCCGCCTTCAACA GTG
3D-A122T-U	CTGGCCTCCCCTGGACCCTCCAGGGGAAACGCC	Production of the A122T mutant
3D-A122T-L	GTTTCCCCTGGAGGGTCCAGGGGAGGCCAGGCG
3D-M296I-U	CTGTTGAAGGCGGGATACCGTCTGGTTGTTCCGCG	Production of the M296I mutant
3D-M296I-L	GAACAACCAGACGGTATCCCGCCTTCAACAGTG
3D-R440W-U	GACCTGATGAGTACTGGCGTCTCTTTGAGCCTTTC	Production of the R440W mutant
3D-R440W-L	AAAGGCTCAAAGAGACGCCAGTACTCATCAGGTCC
3D-T64I-U	TAAACACAAGGAAAACATCAAGATGTCTGAGGAGG	Production of the T64I mutant
3D-T64I-L	CTCCTCAGACATCTTGATGTTTTCCTTGTGTTTAG
3D-D133N-U	CCGTGGTGCGCTCATTAACTTCGAGAACGGCACTG	Production of the D133N mutant
3D-D133N-L	GACAGTGCCGTTCTCGAAGTTAATGAGCGCACCAC