Table 1. Analysis of the protospacers.
| Locus 1 | Locus 2 | |
|---|---|---|
| Protospacer location | ||
| Total | 52 | 108 |
| Plasmid | 50 (96.1%) | 99 (91.7%) |
| Genomic DNA | 2 (3.9%) | 9 (8.3%) |
| Forward strand | 28 (53.8%) | 60 (55.6%) |
| Reverse strand | 24 (46.2%) | 48 (44.4%) |
| Within protein gene | 40 (76.9%) | 72 (66.7%) |
| Intergenic | 12 (23.1%) | 36 (33.3%) |
| PAM sequences | ||
| CCN | 35 (67.3%) | 80 (74.1%) |
| CCA | 12 (23.1%) | 35 (32.4%) |
| CCT | 12 (23.1%) | 20 (18.5%) |
| CCG | 6 (11.5%) | 15 (13.9%) |
| CCC | 5 (9.6%) | 10 (9.3%) |
| No PAM | 17 (32.7%) | 28 (25.9%) |
| Single and multiple spacer insertion | ||
| Single | 42 | 84 |
| Two | 3 | 12 |
| Four | 1 | |
A total of 160 unique spacers inserted into loci 1 and 2 were sequenced and analyzed. The numbers of protospacers matching the expression plasmid and genomic DNA, as well as the protospacers carrying PAM sequences (5′-PAM-protospacer-3′), are given. Percentages are included in brackets.