5T2H
Engineered variant of I-OnuI meganuclease targeting the Human TCRa gene; harbors 43 point mutations relative to wild-type I-OnuI
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ALS BEAMLINE 5.0.1 |
| Synchrotron site | ALS |
| Beamline | 5.0.1 |
| Temperature [K] | 108 |
| Detector technology | CCD |
| Collection date | 2016-04-07 |
| Detector | ADSC QUANTUM 315r |
| Wavelength(s) | 0.976 |
| Spacegroup name | P 21 21 21 |
| Unit cell lengths | 39.696, 68.928, 157.530 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 41.770 - 2.517 |
| R-factor | 0.2171 |
| Rwork | 0.212 |
| R-free | 0.26530 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 3qqy |
| RMSD bond length | 0.003 |
| RMSD bond angle | 0.517 |
| 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 | 2.610 |
| High resolution limit [Å] | 2.520 | 5.430 | 2.520 |
| Rmerge | 0.102 | 0.050 | 0.805 |
| Rmeas | 0.106 | 0.053 | 0.843 |
| Rpim | 0.029 | 0.015 | 0.243 |
| Total number of observations | 198070 | ||
| Number of reflections | 15233 | ||
| <I/σ(I)> | 7.3 | ||
| Completeness [%] | 99.2 | 99.9 | 95.3 |
| Redundancy | 13 | 12.9 | 10.7 |
| CC(1/2) | 0.999 | 0.853 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 6.5 | 293 | 35% (v/v) Pentaerythritol ethoxylate (15/4 EO/OH), 50mM BIS-TRIS pH 6.5, 50mM ammonium sulfate |
