Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli K-12) | MC4100 | Casadaban, 1976 | F- araD139 DlacU169 rpsL150 thi rbsR | |
| Strain, strain background (Escherichia coli K-12) | DRH1119 | This paper | MC4100 ΔsecA::KanR λatt::placUV5-secA | |
| Strain, strain background (Escherichia coli K-12) | DRH1120 | This paper | MC4100 ΔsecA::KanR λatt::placUV5-secAΔMBD | |
| Strain, strain background (Escherichia coli K-12) | DRH1121 | This paper | MC4100 ΔsecA::KanR λatt::placUV5-secAΔCTT | |
| Strain, strain background (Escherichia coli K-12) | DRH663 | This paper | MC4100 ΔsecA::KanR + pDH663 | |
| Strain, strain background (Escherichia coli B) | BL21(DE3) | Lab stock | Escherichia coli Genetic Stock Center (CGSC), Yale University, USA. CGSC#: 12504 http://cgsc2.biology.yale.edu/Strain.php?ID=139459 | MC4100 ΔsecA::KanR + pDH663 |
| Strain, strain background (Escherichia coli B) | DRH584 | This paper | BL21(DE3) containing plasmid pDH584 | |
| Strain, strain background (Escherichia coli B) | DRH1166 | This paper | BL21(DE3) containing plasmid pDH1166 and pSup-Bpa-6TRN | |
| Genetic reagent (phage λ) | lambda InCh | Boyd et al., 2000 | ||
| Antibody | Rabbit anti-SecA antiserum | Huber et al., 2011 | (1:20000) | |
| Antibody | Sheep anti-uL23 antiserum | other | Gift from R. Brimacombe (1:2500) | |
| Antibody | Rabbit anti-thioredoxin-1 antiserum | Sigma-Aldrich (St. Louis, MO, USA) | Catalogue number: T0803 | (1:10000) |
| Antibody | Goat IR700-labelled anti-rabbit | Rockland (Philadelphia, PA, USA) | Catalogue number: 611-130-122 | Discontinued (1:5000) |
| Antibody | Goat IR800-labelled anti-sheep | Rockland (Philadelphia, PA, USA) | Catalogue number: 613-445-002 | (1:10000) |
| Antibody | Donkey HRP-labelled anti-rabbit | GE Healthcare | Catalogue number: NA934V | (1:10000) |
| Recombinant DNA reagent | pCA528 | Andréasson et al., 2010 | pET24 expression vector containing gene encoding His-tagged SUMO protein | |
| Recombinant DNA reagent | pCA597 | Andréasson et al., 2010 | pET24 expression vector containing gene encoding Strep-tagged SUMO protein | |
| Recombinant DNA reagent | pDH543 | This paper | pCA597 containing portion of secA gene corresponding to amino acids829–901 | |
| Recombinant DNA reagent | pDH934 | This paper | pCA597 containing portion of secA gene corresponding to amino acids 875–901 | |
| Recombinant DNA reagent | pDH625 | Huber et al., 2011 | pCA528 containg the secA gene | |
| Recombinant DNA reagent | pDH584 | Huber et al., 2011 | pCA528 containg the secA-biotin gene | |
| Recombinant DNA reagent | pDH1166 | This paper | pDH584 containing amber codon at position corresponding to amino acid 852 in SecA. | |
| Recombinant DNA reagent | pDSW204 | Weiss et al., 1999 | pTrc99a-derived plasmid containing partially disabled trc promoter | |
| Recombinant DNA reagent | pDH692 | Huber et al., 2011 | pDSW204-derived plasmid producing SecA under control of an IPTG-inducible promoter | |
| Recombinant DNA reagent | pDH939 | This paper | pDSW204-derived plasmid producing SecAΔMBD under control of an IPTG-inducible promoter | |
| Recombinant DNA reagent | pDH663 | Huber et al., 2011 | pTrc99b-derived plasmid producing SecA under control of an IPTG-inducible promoter and containing a Spectinomycin resistance gene in place of the bla (ampicillin resistance) gene | |
| Recombinant DNA reagent | pDH787 | Huber et al., 2017 | pCA597 containing full-length secM gene | |
| Recombinant DNA reagent | pDH784 | Huber et al., 2017 | pCA597 containing secM with internal deletion between region encoding signal sequence and the translation arrest sequence | |
| Recombinant DNA reagent | pDH894 | Huber et al., 2017 | pCA528 containing gene encoding malE gene that is translationally fused to sequence encoding the SecM translation arrest sequence | |
| Recombinant DNA reagent | pSup-Bpa-6TRN | Chin et al., 2002 | Plasmid producing orthologous tRNA and tRNA synthetase required for incorporation of Bpa | |
| Peptide, recombinant protein | VipB peptide | Pietrosiuk et al., 2011 | ||
| Peptide, recombinant protein | HRP-coupled Streptactin | IBA Life Sciences (Goettingen, Germany) | Catalogue number: 2-1502-001 | |
| Chemical compound | 4-acetamido-4′-maleimidylstilbene-2,2′-disulfonic acid (AMS) | Invitrogen (Carlsbad, California) | Catalogue number: A485 | |
| Chemical compound, drug | Hydrophilic streptavidin magnetic beads | New England Biolabs (Ipswich, Massachusetts) | Catalogue number: S1421S | |
| Chemical compound, drug | Benzophenylalanine (Bpa) | Bachem (Santa Cruz, CA, USA) | H-p-Bz-Phe-OH Article number:4017646.0005 | |
| Chemical compound, drug | 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) | ThermoScientific Pierce | Catalogue number: 22980 | |
| Chemical compound, drug | Ru(bpy)2(dcbpy) | Sigma Aldrich (St Louis, MO, USA) | Product 96632 | Discontinued |
| Software, algorithm | ATSAS v2.8.3 | European Molecular Biology Laboratory (EMBL) Hamburg | https://www.embl-hamburg.de/biosaxs/software.html | |
| Software, algorithm | PyMol v1.8.0.5 | Schrödinger Scientific | https://pymol.org/2/ | |
| Software, algorithm | GROMACS | Schrödinger Scientific | Pronk et al., 2013 | |
| Chemical compound, drug | PEG/Ion Screen 2 #39 | Hampton Research (Aliso Viejo, CA, USA) | Product HR2-126 | |
| Chemical compound, drug | Morpheus | Molecular Dimensions (Newmarket, Suffolk, UK) | Product MD1-46 | |
| Other | Superose 610/300 GL column | GE Healthcare | Product 17517201 | Discontinued |