Table 2.
DNA primers used in this study.
| Primer name | Nucleotide sequencea |
|---|---|
| angN-F | 5′-ACGACGATTGATGGGTGTAGC-3′ |
| angN-BamHIR | 5′-TTGTATTCACTATGGATCCTTGC-3′ |
| D133A-F | 5′-CGCTTACATATTGATAGCGCTATGATTGCTATTGACCCAG-3′ |
| D133A-R | 5′-CTGGGTCAATAGCAATCATAGCGCTATCAATATGTAAGCG-3′ |
| D138A-F | 5′-CGATATGATTGCTATTGCCCCAGATAGTTGCCGAG-3′ |
| D138A-R | 5′-CTCGGCAACTATCTGGGGCAATAGCAATCATATCG-3′ |
| D575A-F | 5′-GTATTTTCTCGCTTTGCTGCATTAATTCTTGATGCTCGCTC-3′ |
| D575A-R | 5′-GAGCGAGCATCAAGAATTAATGCAGCAAAGCGAGAAAATAC-3′ |
| D580A-F | 5′-CTTTGATGCATTAATTCTTGCTGCTCGCTCCATTGCTTC-3′ |
| D580A-R | 5′-GAAGCAATGGAGCGAGCAGCAAGAATTAATGCATCAAAG-3′ |
| PangN-XhoI | 5′-CTCGAGGTTTCCTCATCAATCCAAAAGGTC-3′ |
| PangN-PstI | 5′-CTGCAGCAACATGCAGCCTGCATTGGTGTTAATTC-3′ |
| PEX-angN | 5′-TGCCCATTATCTTTTCTTCC-3′ |
| angN-T7L | 5′-CGAAAGTTCTATTATTGATGTT-3′ |
| angN-T7R | 5′-ggatcctaatacgactcactatagggaggAGGTAATAAACTAACGGAGAAT-3′ |
a
Restriction sites are underlined, base changed from the wt sequence are shown in italics, T7 promoter sequence in lower case