Table 1.
Sequences of functional 970 loop mutants.
| Sequencesb |
Number of mutationsc | Percent functiond | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Samples | 964 | 965 | 966 | 967 | 968 | 969 | 970 | 971 | ||
| WTa | A | U | G | C | A | A | C | G | 0 | 100 |
| 1 | A | U | G | A | A | A | C | G | 1 | 119.8 (±1.6) |
| 2 | A | U | G | C | A | A | C | U | 1 | 94.3 (±3.3) |
| 3 | A | C | G | A | U | A | C | G | 3 | 54.3 (±2.8) |
| 4 | U | C | G | C | U | A | A | C | 5 | 39.8 (±2.6) |
| 5 | A | U | G | C | U | A | A | C | 3 | 39.7 (±2.0) |
| 6 | A | U | G | A | U | A | C | G | 2 | 39.2 (±1.2) |
| 7 | A | U | G | C | G | G | C | G | 2 | 38.1 (±1.3) |
| 8 | A | U | C | A | C | A | C | G | 3 | 37.3 (±1.6) |
| 9 | A | C | G | A | A | A | C | A | 3 | 35.1 (±1.7) |
| 10 | A | U | A | A | U | G | C | G | 4 | 34.9 (±1.7) |
| 11 | A | U | G | C | C | A | C | A | 2 | 34.4 (±2.5) |
| 12 | A | G | G | G | A | G | A | A | 5 | 34.3 (±1.9) |
| 13 | A | U | G | C | C | A | C | C | 2 | 34.3 (±1.3) |
| 14 | A | U | C | G | A | G | G | G | 4 | 34.2 (±0.5) |
| 15 | A | C | G | C | G | A | C | G | 2 | 33.7 (±1.0) |
| 16 | A | U | A | A | U | G | C | C | 5 | 32.2 (±0.4) |
| 17 | A | C | G | C | C | G | C | G | 3 | 32.0 (±1.9) |
| 18 | A | A | G | C | G | G | C | G | 3 | 31.6 (±1.2) |
| 19 | A | A | A | C | U | A | C | G | 3 | 31.5 (±1.3) |
| 20 | A | C | A | A | U | A | C | C | 5 | 31.4 (±1.8) |
| 21 | U | C | G | U | U | A | C | A | 5 | 30.7 (±0.4) |
| 22 | U | C | G | C | U | A | C | C | 4 | 30.0 (±0.8) |
| 23 | A | C | G | A | U | A | C | U | 4 | 29.8 (±1.6) |
| 24 | A | C | A | A | U | A | C | C | 5 | 29.7 (±1.8) |
| 25 | A | A | G | G | U | G | C | G | 4 | 29.7 (±1.3) |
| 26 | A | A | G | U | U | A | C | C | 4 | 28.9 (±2.1) |
| 27 | A | C | G | G | U | A | C | C | 4 | 28.7 (±3.2) |
| 28 | A | U | A | A | U | A | C | A | 4 | 28.5 (±1.8) |
| 29 | A | C | A | A | C | A | C | G | 4 | 28.3 (±2.6) |
| 30 | A | C | A | A | U | A | A | G | 5 | 28.1 (±1.4) |
| 31 | A | C | A | G | U | A | A | G | 5 | 27.6 (±1.5) |
| 32 | U | A | G | C | U | A | C | A | 4 | 27.3 (±1.5) |
| 33 | A | G | A | G | U | A | C | G | 4 | 27.1 (±2.1) |
| 34 | C | A | C | A | C | A | C | A | 6 | 26.9 (±0.7) |
| 35 | A | U | A | G | U | A | A | G | 4 | 26.8 (±1.5) |
| 36 | A | C | G | C | U | G | C | C | 4 | 26.7 (±1.7) |
| 37 | A | C | A | C | C | A | C | A | 4 | 26.5 (±2.4) |
| 38 | U | C | G | C | U | A | A | G | 4 | 26.4 (±1.3) |
| 39 | A | U | G | C | U | A | A | A | 3 | 26.3 (±3.1) |
| 40 | A | U | A | A | U | A | A | G | 4 | 26.1 (±2.7) |
| 41 | A | C | G | A | U | G | A | G | 5 | 26.1 (±1.3) |
| 42 | A | U | A | C | C | A | C | G | 2 | 26.0 (±2.3) |
| 43 | A | C | A | C | G | A | C | G | 3 | 25.8 (±0.8) |
| 44 | A | A | G | C | U | A | A | A | 4 | 25.7 (±0.1) |
| 45 | A | C | A | A | U | A | C | U | 5 | 25.0 (±0.8) |
| 46 | A | C | A | A | U | A | A | C | 6 | 24.7 (±2.7) |
| 47 | A | U | G | C | C | G | C | A | 3 | 24.5 (±1.3) |
| 48 | C | A | A | A | U | A | C | A | 6 | 24.5 (±0.6) |
| 49 | A | A | A | C | U | A | C | A | 4 | 24.5 (±2.0) |
| 50 | A | C | A | C | U | A | A | C | 5 | 24.4 (±1.2) |
| 51 | A | U | A | A | C | A | C | C | 4 | 23.8 (±2.4) |
| 52 | A | G | G | C | U | A | A | C | 4 | 23.6 (±0.6) |
| 53 | A | C | U | C | A | A | C | G | 2 | 23.4 (±0.1) |
| 54 | C | C | G | U | U | A | C | A | 5 | 23.4 (±1.0) |
| 55 | U | C | A | A | U | A | A | C | 7 | 23.1 (±1.4) |
| 56 | A | C | A | G | U | A | A | C | 6 | 23.0 (±1.4) |
| 57 | A | A | U | C | C | A | C | G | 3 | 22.6 (±0.6) |
| 58 | A | C | A | C | U | G | C | C | 5 | 22.4 (±1.7) |
| 59 | A | U | A | A | U | A | A | C | 5 | 22.0 (±2.6) |
| 60 | A | U | G | G | C | G | C | A | 4 | 21.6 (±0.5) |
| 61 | A | A | G | C | G | A | A | C | 4 | 21.6 (±2.0) |
| 62 | A | C | A | A | C | A | C | C | 5 | 21.1 (±2.0) |
| 63 | A | A | G | C | A | G | U | G | 3 | 20.9 (±1.4) |
| 64 | A | C | A | A | U | G | A | C | 7 | 20.8 (±0.4) |
| 65 | A | U | A | C | G | A | C | G | 2 | 20.5 (±0.4) |
| 66 | A | C | A | A | C | C | C | G | 5 | 19.9 (±2.9) |
| 67 | A | U | G | A | U | G | A | C | 5 | 19.7 (±2.0) |
| 68 | A | C | A | A | U | A | A | A | 6 | 19.2 (±1.5) |
| 69 | A | A | G | G | A | G | G | U | 5 | 19.2 (±1.9) |
| 70 | A | C | A | C | A | A | A | G | 3 | 18.8 (±1.1) |
| 71 | A | U | A | C | U | A | A | A | 4 | 18.3 (±1.6) |
| 72 | A | A | G | C | U | G | U | C | 5 | 18.1 (±0.6) |
| 73 | A | U | A | A | C | A | A | A | 5 | 18.1 (±1.1) |
| 74 | A | U | A | A | U | C | C | C | 5 | 18.0 (±0.2) |
| 75 | C | A | C | A | U | A | C | G | 5 | 17.7 (±2.0) |
| 76 | A | U | G | C | A | A | G | A | 2 | 17.6 (±2.2) |
| 77 | A | C | C | C | U | A | C | G | 3 | 17.5 (±1.3) |
| 78 | A | C | G | C | A | A | U | G | 2 | 16.9 (±0.2) |
| 79 | A | U | C | A | A | G | C | G | 3 | 16.2 (±1.0) |
| 80 | A | C | A | A | A | G | C | C | 5 | 16.1 (±2.2) |
| 81 | A | U | A | U | U | C | C | A | 5 | 15.8 (±1.2) |
| 82 | A | C | A | A | A | A | C | A | 4 | 15.0 (±2.0) |
| 83 | A | U | A | G | C | A | A | G | 4 | 15.0 (±2.6) |
| 84 | A | U | A | U | C | G | A | G | 5 | 14.8 (±0.4) |
| 85 | A | C | A | U | C | A | A | C | 6 | 13.9 (±1.2) |
| 86 | A | U | A | C | U | G | U | G | 4 | 13.6 (±1.3) |
| 87 | A | C | A | A | A | G | A | A | 6 | 13.6 (±0.8) |
| 88 | A | C | G | C | U | G | C | G | 3 | 12.9 (±0.8) |
| 89 | A | U | U | A | C | G | C | G | 4 | 12.5 (±0.1) |
| 90 | C | A | C | U | C | A | C | G | 5 | 12.4 (±0.3) |
| 91 | A | C | A | U | C | A | C | U | 5 | 12.3 (±0.1) |
| 92 | A | C | A | G | U | G | A | A | 7 | 12.2 (±2.8) |
| 93 | A | C | C | C | U | G | C | A | 5 | 11.8 (±1.4) |
| 94 | A | A | A | C | A | G | C | A | 4 | 11.5 (±3.4) |
| 95 | A | U | C | A | G | C | U | A | 6 | 11.4 (±2.8) |
| 96 | C | A | C | A | C | G | A | G | 7 | 11.4 (±2.6) |
| 97 | A | U | U | C | U | A | C | C | 3 | 11.4 (±2.0) |
| 98 | A | C | U | A | U | U | C | G | 5 | 11.0 (±0.2) |
| 99 | U | C | U | C | U | A | A | C | 6 | 10.9 (±1.0) |
| 100 | A | U | G | A | A | G | C | C | 3 | 10.4 (±3.4) |
| 101 | U | C | G | G | C | C | G | A | 7 | 10.2 (±3.2) |
| 102 | C | A | C | U | U | G | C | A | 7 | 10.1 (±0.4) |
| 103 | A | C | U | A | U | G | C | C | 6 | 10.0 (±2.8) |
| 104 | A | C | C | C | U | A | C | C | 4 | 10.0 (±1.9) |
| 105 | A | U | C | G | U | G | C | G | 4 | 9.8 (±3.0) |
| 106 | A | C | C | U | U | A | C | C | 5 | 9.5 (±1.8) |
| 107 | A | A | A | A | U | G | A | C | 7 | 9.5 (±1.3) |
a
Sequence and function of the unmutated construct (WT, wild type).
b
The 970 loop sequences selected from the pool of functional mutants. Mutations are underlined.
c
The number of mutations in each sequence.
d
The level of function for each mutant with standard error. The function is an based on at least three independent assays.