8DH2

T7 RNA polymerase elongation complex with unnatural base dDs-ATP mismatch

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]	100
Detector technology	CCD
Collection date	2022-04-21
Detector	ADSC QUANTUM 315r
Wavelength(s)	1
Spacegroup name	P 1
Unit cell lengths	78.702, 85.047, 200.477
Unit cell angles	89.87, 85.38, 69.51

Refinement procedure

Resolution	47.370 - 2.900
R-factor	0.2609
Rwork	0.260
R-free	0.30750
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	8dh0
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHENIX (1.19)
Refinement software	PHENIX (1.19)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.550	48.550	2.940
High resolution limit [Å]	2.890	15.830	2.890
Rmerge	0.139	0.022	2.003
Rmeas	0.164	0.025	2.370
Rpim	0.087	0.013	1.251
Total number of observations	376957	2336	18952
Number of reflections	106891	632	5354
<I/σ(I)>	5.5	31.8	0.5
Completeness [%]	98.3	95.5	98
Redundancy	3.5	3.7	3.5
CC(1/2)	0.995	0.999	0.273

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.1	298	10% PEG 8000, 8% glycerol, 5 mM B-mercaptoethanol, 100 mM Tris pH 8.1