8DH4

T7 RNA polymerase elongation complex with unnatural base dPa-DsTP pair

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.1
Synchrotron site	ALS
Beamline	8.2.1
Temperature [K]	100
Detector technology	CCD
Collection date	2021-12-09
Detector	ADSC QUANTUM 315r
Wavelength(s)	1
Spacegroup name	P 1
Unit cell lengths	78.779, 86.230, 201.463
Unit cell angles	89.85, 85.39, 69.59

Refinement procedure

Resolution	43.200 - 2.800
R-factor	0.2334
Rwork	0.233
R-free	0.27450
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]	48.840	48.840	2.800
High resolution limit [Å]	2.750	15.060	2.750
Rmerge	0.099	0.037	2.239
Rmeas	0.115	0.044	2.595
Rpim	0.059	0.024	1.310
Total number of observations	481832	2503	24587
Number of reflections	125883	668	6284
<I/σ(I)>	8.2	29	0.6
Completeness [%]	97.8	85.4	97.8
Redundancy	3.8	3.7	3.9
CC(1/2)	0.995	0.973	0.253

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