Table 2.
Primers used in this study.
| Primer | Sequence (5′-3′) | Purpose | Source or reference |
|---|---|---|---|
| P001-Δcas1-up-F | ATAAAAATCCGATATGGCTCAACAAAGAAAAAGG | RA-CH-2 Δcas1, Δcas1-cas2 | This study |
| P002-Δcas1-up-R | TGTATTCACGAAAATCCATTATTTTTAATATTCTCC | This study | |
| P003-Δcas1-spc-F | AATAATGGATTTTCGTGAATACATGTTATAATAACT | This study | |
| P004-Δcas1-spc-R | TTAAACTTCATGCTTACCAATTAGAATGAATA | RA-CH-2 Δcas1 | This study |
| P005-Δcas1-down-F | CTAATTGGTAAGCATGAAGTTTAATCGTTTTAATG | This study | |
| P006-Δcas1-down-R | CACTCGCAATCTGCTGTAAATCGTTTTG | This study | |
| P007-Δcas2-up-F | CCTTTGTATGGGCACTCCGAATAT | RA-CH-2 Δcas2 | This study |
| P008-Δcas2-up-R | GTATTCACGAATTAAACTTCATAAAACTCAGGC | This study | |
| P009-Δcas2-spc-F | TGAAGTTTAATTCGTGAATACATGTTATAATAACTA | This study | |
| P010-Δcas2-spc-R | CTTTTTCTATTCGCTTACCAATTAGAATGAATAT | RA-CH-2 Δcas2, Δcas1-cas2 | This study |
| P011-Δcas2-down-F | TAATTGGTAAGCGAATAGAAAAAGACTTTACATC | This study | |
| P012-Δcas2-down-R | GCAACATAGACGAGATTGAGATATGACAACAC | This study | |
| P013-Δcas1-cas2-Ident-F | AAAGATTGAAAAGGTGGTGG | Identification for all the deletion mutant strains | This study |
| P014-Δcas1-cas2-Ident-R | CGCAACATTTTACACGAAG | This study | |
| P015-cas1-exp-F | CGGAATTCATATGCATCATCATCATCATCACATGCTTT ACCGTTCTATCTAT |
pET30a::cas1, pET30a::cas1E149A, pET30a::cas1H206A, pET30a::cas1E221A, pET30a::cas1-cas2 | This study |
| P016-cas1-exp-R | CCGCTCGAGTTAAACTTCATAAAACTCAGG | pET30a::cas1, pET30a::cas1E149A, pET30a::cas1H206A, pET30a::cas1E221A | This study |
| P017-cas2-exp-R | CCGCTCGAGCTACTTATCGTCGTCATCCTTGTAATC AAAAAGTTCTAATTGC |
pET30a::cas1-cas2 | This study |
| P018-CRISPR-F | AAACCAACCTATCAGCAA | pGEM::CRISPR | This study |
| P019-CRISPR-R | AATGGATTTTGGGAGTAT | This study | |
| P020-E149A-R | TGCAATGCCTGCCATATTGGTG | pET30a::cas1E149A | This study |
| P021-E149A-F | CACCAATATGGCAGGCATTGCA | This study | |
| P022-H206A-R | ATTTATTCTTAGCAAAAATCCCCAATACAGGC | pET30a::cas1H206A | This study |
| P023-H206A-F | CCTGTATTGGGGATTTTTGCTAAGAATAAAT | This study | |
| P024-E221A-R | ACGGTAAGGTGCCATTAAATCGTCT | pET30a::cas1E221A | This study |
| P025-E221A-F | ACGATTTAATGGCACCTTACCGT | This study |