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