5CD4

The Type IE CRISPR Cascade complex from E. coli, with two assemblies in the asymmetric unit arranged back-to-back

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	80
Detector technology	IMAGE PLATE
Collection date	2014-03-22
Detector	MAR scanner 300 mm plate
Wavelength(s)	1.0
Spacegroup name	P 21 21 21
Unit cell lengths	105.990, 244.800, 426.740
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.735 - 3.200
R-factor	0.2127
Rwork	0.212
R-free	0.24960
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1VY8
RMSD bond length	0.004
RMSD bond angle	0.740
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.10pre_2089: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.740	3.250
High resolution limit [Å]	3.200	3.200
Number of reflections	183649
<I/σ(I)>	8	2
Completeness [%]	100.0	100
Redundancy	7.3	6.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		291	0.1 M HEPES pH 7.0, 0-0.1 M KCl, and 8-14 percent (w/v) PEG 8000. The crystal used in this study grew in the presence of an 11-nt ssDNA containing a 5-CTT-3 PAM and nine nucleotides complementary to the crRNA-guide sequence in a 2:1 oligonucleotide:protein ratio. However, the target DNA is not observed in the electron