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- PDB-7rcd: Second stage reengineered variant of I-OnuI targeting human PD1 g... -
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Open data
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Basic information
Entry | Database: PDB / ID: 7rcd | ||||||||||||
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Title | Second stage reengineered variant of I-OnuI targeting human PD1 gene with activity enhancing substitutions | ||||||||||||
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![]() | DNA BINDING PROTEIN/DNA / meganuclease / gene editing / protein engineering / DNA BINDING PROTEIN-DNA complex | ||||||||||||
Function / homology | DNA / DNA (> 10)![]() | ||||||||||||
Biological species | Synthetic construct (others) | ||||||||||||
Method | ![]() ![]() ![]() ![]() | ||||||||||||
![]() | Ubilla-Rodriguez, N.C. / Werther, R.A. / Stoddard, B.L. | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Characterization of the stepwise engineering and optimization of a retargeted DNA binding protein and gene-editing meganuclease Authors: Werther, R.A. / Ubilla-Rodriguez, N.C. / Smiley, A.T. / Havens, K. / Lambert, A.R. / Stoddard, B.L. | ||||||||||||
History |
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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Download
PDBx/mmCIF format | ![]() | 103.1 KB | Display | ![]() |
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PDB format | ![]() | 70.6 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
Others | ![]() |
-Validation report
Summary document | ![]() | 439.2 KB | Display | ![]() |
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Full document | ![]() | 441.3 KB | Display | |
Data in XML | ![]() | 13.4 KB | Display | |
Data in CIF | ![]() | 17.3 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7rccC ![]() 7rceC ![]() 7rcfC ![]() 7rcgC ![]() 3qqyS S: Starting model for refinement C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
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Assembly
Deposited unit | ![]()
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Unit cell |
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Components
#1: Protein | Mass: 34993.363 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Synthetic construct (others) / Production host: ![]() ![]() | ||||||
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#2: DNA chain | Mass: 8064.180 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Synthetic construct (others) | ||||||
#3: DNA chain | Mass: 7917.066 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Synthetic construct (others) | ||||||
#4: Chemical | #5: Water | ChemComp-HOH / | Has ligand of interest | N | Sequence details | The wild type homing endonuclease was extensively engineered to generate I-OnuI_e-hPD1-c. | |
-Experimental details
-Experiment
Experiment | Method: ![]() |
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Sample preparation
Crystal | Density Matthews: 2.24 Å3/Da / Density % sol: 44.97 % |
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Crystal grow | Temperature: 293 K / Method: vapor diffusion, hanging drop / pH: 7 Details: 100mM HEPES pH 7.0, 200mM ammonium sulfate, 25% PEG 3350 |
-Data collection
Diffraction | Mean temperature: 108 K / Serial crystal experiment: N | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Diffraction source | Source: ![]() ![]() ![]() | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Detector | Type: DECTRIS PILATUS 6M / Detector: PIXEL / Date: Aug 29, 2019 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Radiation | Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Radiation wavelength | Wavelength: 1 Å / Relative weight: 1 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Reflection | Resolution: 2.448→50 Å / Num. obs: 17555 / % possible obs: 99.8 % / Redundancy: 5.7 % / Biso Wilson estimate: 54.61 Å2 / Rmerge(I) obs: 0.038 / Rpim(I) all: 0.018 / Rrim(I) all: 0.043 / Χ2: 1.032 / Net I/σ(I): 23.3 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Reflection shell | Diffraction-ID: 1
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-Phasing
Phasing | Method: ![]() | |||||||||
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Phasing MR |
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Processing
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Refinement | Method to determine structure: ![]() Starting model: 3qqy Resolution: 2.448→42.478 Å / SU ML: 0.36 / Cross valid method: THROUGHOUT / σ(F): 1.37 / Phase error: 30.84 / Stereochemistry target values: ML
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Solvent computation | Shrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Displacement parameters | Biso max: 117.97 Å2 / Biso mean: 62.9953 Å2 / Biso min: 28.57 Å2 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refinement step | Cycle: final / Resolution: 2.448→42.478 Å
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Refine LS restraints |
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LS refinement shell | Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0
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