5T2O
Engineered variant of I-OnuI meganuclease targeting the Anopheles AGAP011377 gene; harbors 53 point mutations relative to wild-type I-OnuI
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ALS BEAMLINE 5.0.3 |
| Synchrotron site | ALS |
| Beamline | 5.0.3 |
| Temperature [K] | 108 |
| Detector technology | CCD |
| Collection date | 2016-01-30 |
| Detector | ADSC QUANTUM 315r |
| Wavelength(s) | 0.976 |
| Spacegroup name | P 21 21 21 |
| Unit cell lengths | 40.180, 64.835, 167.005 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 42.236 - 2.801 |
| R-factor | 0.2298 |
| Rwork | 0.224 |
| R-free | 0.28080 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 3qqy |
| RMSD bond length | 0.003 |
| RMSD bond angle | 0.545 |
| Data reduction software | HKL-2000 |
| Data scaling software | HKL-2000 |
| Phasing software | PHASER (2.5.6) |
| Refinement software | PHENIX (1.9_1692) |
Data quality characteristics
| Overall | Inner shell | Outer shell | |
| Low resolution limit [Å] | 50.000 | 50.000 | 2.900 |
| High resolution limit [Å] | 2.800 | 6.030 | 2.800 |
| Rmerge | 0.155 | 0.060 | 0.877 |
| Rmeas | 0.161 | 0.062 | 0.920 |
| Rpim | 0.045 | 0.017 | 0.273 |
| Total number of observations | 140749 | ||
| Number of reflections | 11189 | ||
| <I/σ(I)> | 4.5 | ||
| Completeness [%] | 98.4 | 100 | 96.9 |
| Redundancy | 12.6 | 12.6 | 10.9 |
| CC(1/2) | 0.999 | 0.859 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 7.5 | 293 | 33% (v/v) pentaerythritol ethoxylate (15/4 EO/OH), 50mM HEPES pH 7.5, 50mM ammonium sulfate |
