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. 2025 Mar 13;44(9):2501–2513. doi: 10.1038/s44318-025-00408-1

Reagents and tools table

Reagent/resource Reference or source Identifier or catalog number
Experimental models
E. coli BL21 FtsH-FLAG (C-terminal) This work following: Reisch CR and Prather KLJ (2017) Strain AG1_FR1_B1_A1
BL21 FtsH-FLAG (C-terminal) ΔhflK/C This work following: Reisch CR and Prather KLJ (2017) Strain AG2_Fr1_B1_A2
Recombinant DNA
BL21_C-FLAG construct Integrated DNA Technologies, Inc. Sequence provided as source data file BL21_C-flag.dna
pKDsgRNA-p15a Addgene Plasmid #62656
pCas9-CR4 Addgene Plasmid #62655
GTGAAAGCACTGATTGAGCGTAACTATAATCGTGCGCGTCAGCTTCTGACCGACAATATGGATATTCTGCATGCGATGAAAGATGCTCTCATGAAATATGAGACTATCGACGCACCGCAGATTGATGACCTGATGGCACGTCGCGATGTACGTCCGCCAGCGGGCTGGGAAGAACCAGGCGCTTCTAACAATTCTGGCGACAATGGTAGTCCAAAGGCTCCTCGTCCGGTTGATGAACCGCGTACGCCGAACCCGGGTAACACGATGTCAGAGCAGTTAGGCGACAAGAGCGGGTCAGGAGACTATAAGGACGACGACGACAAGTAA Integrated DNA Technologies, Inc. Geneblock sequence containing FtsH Flag Tag (shown underline) and protospacer (shown in bold)
Antibodies
NA NA NA
Oligonucleotides and other sequence-based reagents
Recombineering oligos Integrated DNA Technologies, Inc. NA
Chemicals, enzymes, and other reagents
Pyruvate Kinase/Lactic Dehydrogenase enzymes from rabbit muscle Sigma Aldrich, Inc. P0294
NADH, grade II Roche, Inc. CAS# 606688
Phosphoenolpyruvate Roche, Inc. 10108294001
Carboxy-DIBMA Dr. Sandro Keller’s lab University of Graz Described in this work
Graphene-supported grids Grassetti et al, 2023 200-mesh Quantifoil 2/1 copper grids, pre-treated with UV/ozone for 7 min using a Bioforce UV/ozone cleaner
1-myristoyl-2-{6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl}-sn-glycero-3-phosphocholine Avanti Inc. 810122
POPC Avanti Inc. 850457
POPE Avanti Inc. 850757
NBD-PE Avanti Inc. 810153C
DDM Anatrace Inc. D310
GDN Anatrace Inc. GDN101
Bio-Beads SM-2 Adsorbents #1523920 Bio-Rad Inc. #1523920
S-Trap Micro Columns Protifi inc. C02-micro-80
Software
CryoSPARC Punjani et al, 2017 V3.0; v4.0
ChimeraX Pettersen et al, 2021 V1.2
Coot Casanal et al, 2020 V0.9.4
Phenix Liebschner et al, 2019 V1.14
cryoDRGN Zhong et al, 2021 V0.3.5
Warp Tegunov and Cramer, 2019 V1.0.9
IMOD Mastronarde and Held, 2017 V4.11.12
RELION Zivanov et al, 2018 V3.1.3
M Tegunov et al, 2021 V1.0.9
tomoDRGN Powell and Davis, 2024 V0.2.2
Spectronaut Biognosys AG V15
Other
Titan Krios G3i – Gatan BioQuantum K3 Thermo Fisher and Gatan n/a
Q-exactive HF-X Thermo Fisher n/a