Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | BL21 Star (DE3) | Thermo Fisher Scientific | C6010-03 | |
| Strain, strain background (Escherichia coli) | BL21 (DE3) | New England Biolabs | C2527I | |
| Recombinant DNA reagent | pET52b | EMD Millipore | 72554 | |
| Recombinant DNA reagent | pET52b/ His6 Cas4 | Lee et al., 2018 | N/A | |
| Recombinant DNA reagent | pSV272/ His6-MBP-TEV Cas1 | Lee et al., 2018 | N/A | |
| Recombinant DNA reagent | pSV272/ His6-MBP-TEV Cas2 | Lee et al., 2018 | N/A | |
| Recombinant DNA reagent | pRKSUF017 | Takahashi and Tokumoto, 2002 | N/A | |
| Commercial assay or kit | QIAprep Spin Miniprep Kit | Qiagen | #27106 | |
| Commercial assay or kit | Wizard SV Gel and PCR Clean-up system | Promega | #A9282 | |
| Commercial assay or kit | Wizard Plus SV minipreps DNA purification system | Promega | #A1460 | |
| Commercial assay or kit | HisPur Ni-NTA Spin columns | Thermo Fisher Scientific | #88224 | |
| Commercial assay or kit | HisPur Ni-NTA resin | Thermo Fisher Scientific | #88223 | |
| Commercial assay or kit | HiTrap SP HP | GE Healthcare | #7115201 | |
| Commercial assay or kit | HiTrap Heparin HP | GE Healthcare | #17-0407-03 | |
| Commercial assay or kit | HiLoad 16/600 Superdex 200 | GE Healthcare | #28989335 | |
| Commercial assay or kit | HiLoad 16/600 Superdex 75 | GE Healthcare | #28989333 | |
| Commercial assay or kit | Sequenase Version 2.0 DNA Sequencing Kit | Thermo Fisher Scientific | 707701KT | |
| Software, algorithm | Scipion | de la Rosa-Trevín et al., 2016 | scipion.i2pc.es | |
| Software, algorithm | Xmipp | Abrishami et al., 2013; Sorzano et al., 2013; Vargas et al., 2013 | xmipp.i2pc.es | |
| Software, algorithm | RELION | Scheres, 2012 | mrc-lmb.cam.ac.uk/reli | |
| Software, algorithm | RELION | Scheres, 2012 | on/index.php/Main_Page | |
| Software, algorithm | Phyre2 | Kelley et al., 2015 | sbg.bio.ic.ac.uk/phyre2/html/page.cgi?id = index | |
| Software, algorithm | Chimera | Pettersen et al., 2004 | cgl.ucsf.edu/chimera/ | |
| Software, algorithm | Segger | Pintilie et al., 2010 | cryoem.bcm.edu/cryoem/downloads/segger | |
| Other | Formvar/Carbon 400 mesh, Copper approx. grid hole size: 42 μm | Ted Pella, Inc. | 01754 F | |
| Other | Pyrobaculum calidifontis Cas4 | Lemak et al., 2014 | PDB: 4R5Q | Deposited data |
| Other | Archaeoglobus fulgidus Cas1 | Kim et al., 2013 | PDB: 4N06 | Deposited data |
| Other | Bacillus halodurans Cas2 | Nam et al., 2012 | PDB: 4ES3 | Deposited data |
| Other | Escherichia coli Cas1-Cas2 | Nuñez et al., 2014 | PDB: 4P6I | Deposited data |
| Other | Escherichia coli Cas1-Cas2-prespacer | Nuñez et al., 2015b | PDB: 5DS4 | Deposited data |
| Other |
Enterococcus
faecalis Cas1-Cas2-prespacer |
Xiao et al., 2017 | PDB: 5XVN | Deposited data |
| Other |
Enterococcus
faecalis Cas1-Cas2-full site |
Xiao et al., 2017 | PDB: 5XVP | Deposited data |
| Other | Bacillus halodurans Cas1-Cas2-target | This paper | EMDB-20127 | Deposited data |
| Other | Bacillus halodurans Cas4-Cas1-Cas2-target asymmetrical | This paper | EMDB-20128 | Deposited data |
| Other | Bacillus halodurans Cas4-Cas1-Cas2-target symmetrical | This paper | EMDB-20129 | Deposited data |
| Other | Bacillus halodurans Cas4-Cas1-Cas2-prespacer asymmetrical | This paper | EMDB-20130 | Deposited data |
| Other | Bacillus halodurans Cas4-Cas1-Cas2-prespacer symmetrical | This paper | EMDB-20131 | Deposited data |
| Chemical compound, drug | Agar | AMRESCO | #J637-1kg | |
| Chemical compound, drug | Carbenicillin disodium salt | RPI | #C46000-25.0 | |
| Chemical compound, drug | Kanamycin monosulfate | RPI | #K22000-25.0 | |
| Chemical compound, drug | Ampicillin | RPI | #A40040-100.0 | |
| Chemical compound, drug | Tetracycline HCl | RPI | #T17000-25.0 | |
| Chemical compound, drug | LB Broth (Miller) | Thermo Fisher Scientific | #BP1426-2 | |
| Chemical compound, drug | IPTG | RPI | #I56000-100.0 | |
| Chemical compound, drug | DTT | RPI | #D11000-100.0 | |
| Chemical compound, drug | Tryptone | RPI | #T60060-5000.0 | |
| Chemical compound, drug | Sodium chloride | AMRESCO | #7647–14.5 | |
| Chemical compound, drug |
Yeast extract | Thermo Fisher Scientific | #BP1422-2 | |
| Chemical compound, drug | Agarose | Thermo Fisher Scientific | #BP160-500 | |
| Chemical compound, drug | HEPES | Thermo Fisher Scientific | #BP310-1 | |
| Chemical compound, drug | Sodium phosphate dibasic heptahydrate | Thermo Fisher Scientific | #S373-3 | |
| Chemical compound, drug | Glycerol | VWR analytical BDH | #BDH1172-4LP | |
| Chemical compound, drug | Imidazole | Thermo Fisher Scientific | #O31960599 | |
| Chemical compound, drug | PMSF | RPI | #P20270-25.0 | |
| Chemical compound, drug | Ferrous sulfate | Thermo Fisher Scientific | #I146-500 | |
| Chemical compound, drug | Ferric sulfate | Sigma | #F3388-250G | |
| Chemical compound, drug | L-Cysteine free base | MP Biomedicals | #194646 | |
| Chemical compound, drug | Manganese chloride tetrahydrate | Thermo Fisher Scientific | #M87-100 | |
| Chemical compound, drug | Potassium chloride | RPI | #D41000-2500.0 | |
| Chemical compound, drug | Brilliant blue R-250 | RPI | #B43000-50.0 | |
| Chemical compound, drug | 40% Acrylamide/Bis solution, 19:1 | Thermo Fisher Scientific | #BP1406-1 | |
| Chemical compound, drug | Urea | RPI | #U20200-25000.0 | |
| Chemical compound, drug | Boric acid | RPI | #B32050-5000.0 | |
| Chemical compound, drug | Tris | RPI | #T60040-5000.0 | |
| Chemical compound, drug |
EDTA | Thermo Fisher Scientific | #BP120-1 | |
| Chemical compound, drug | 2X RNA loading dye | New England Biolabs | #B0363A | |
| Chemical compound, drug | T4 Polynucleotide Kinase | New England Biolabs | #M0201L | |
| Chemical compound, drug | [γ-32P]-ATP | Perkin Elmer | #BLU502A250UC | |
| Chemical compound, drug |
Phenol:Chloroform:Isoamyl Alcohol (25:24:1) | Thermo Fisher Scientific | #15593049 |