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