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isopropyl β d 1 thiogalactopyranoside iptg  (Apollo Scientific Ltd)

 
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    Apollo Scientific Ltd isopropyl β d 1 thiogalactopyranoside iptg
    Isopropyl β D 1 Thiogalactopyranoside Iptg, supplied by Apollo Scientific Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/iptg/1+d+isopropyl+thiogalactopyranoside+%CE%B2/pm42264237-72-0-6
    Average 86 stars, based on 1 article reviews
    isopropyl β d 1 thiogalactopyranoside iptg - by Bioz Stars, 2026-09
    86/100 stars

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    Recombinant:

    Article Title: INF2 formin variants linked to human inherited kidney disease reprogram the transcriptome, causing mitotic chaos and cell death
    Article Snippet: .. Subsequently, the temperature was lowered to 20 °C, and recombinant protein expression was induced by adding 0.5 mM IPTG (Apollo Scientific, # BIMB1008) for 16 h. Cell lysates from HEK293T cells expressing GFP alone or GFP-centrins 1–3 or GFP-CaM were incubated with assay buffer (20 mM Tris–HCl, pH 7.8, 150 mM NaCl, 1% Nonidet P-40, 1% glycerol, 1 mM sodium orthovanadate, 0.1 mM PMSF, and 5 mM CaCl 2 ), supplemented with a commercial cocktail of protease inhibitors (Merck, # 11697498001) at 4 °C, alongside 30 μg of the indicated GST-fused proteins immobilized on glutathione-Sepharose 4B beads (Merck, # GE17-0756-01). ..

    Article Title: INF2 formin variants linked to human inherited kidney disease reprogram the transcriptome, causing mitotic chaos and cell death.
    Article Snippet: .. Subsequently, the temperature was lowered to 20 °C, and recombinant protein expression was induced by adding 0.5 mM IPTG (Apollo Scientific, # BIMB1008) for 16 h. Cell lysates from HEK293T cells expressing GFP alone or GFP-centrins 1–3 or GFP-CaM were incubated with assay buffer (20 mM Tris–HCl, pH 7.8, 150 mM NaCl, 1% Nonidet P-40, 1% glycerol, 1 mM sodium orthovanadate, 0.1 mM PMSF, and 5 mM CaCl2), supplemented with a commercial cocktail of protease inhibitors (Merck, # 11697498001) at 4 °C, alongside 30 μg of the indicated GST-fused proteins immobilized on glutathioneSepharose 4B beads (Merck, # GE17-0756-01). ..

    Article Title: De novo cysteine biosynthesis in Pseudomonas aeruginosa: characterization of the two main cysteine synthase isoforms
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains See Table 3 for bacterial strains Chemicals, peptides, and recombinant proteins Yeast extract PanReac AppliChem Cat# A1552,1000 Tryptone PanReac AppliChem Cat# A1553,1000 Sodium chloride (for protein expression and purification) PanReac AppliChem Cat# 131659.1211 Sodium chloride (for M9 preparation) Sigma-Aldrich Cat# 746398 Phosphate buffered saline Sigma-Aldrich Cat# P4417-50TAB D-(+)-Glucose Sigma Life Science Cat# G8270-10 KG Isopropil β-D-1-tiogalattopiranoside, IPTG, Isopropil β-D-tiogalattoside (IPTG) Apollo Scientific Cat# BIMB1008 Phenylmethanesulphonyl fluoride (PMSF) Apollo Scientific Cat# PC6222M Benzamidine Fluka Cat# 12072 Pepstatin A PanReac AppliChem Cat# A2205,0010 Pyridoxal-5 ′ -phosphate hydrate (PLP) Sigma-Aldrich Cat# P9255-5G Lysozime from chicken egg white Sigma-Aldrich Cat# 62971-10G-F Ethylenediaminetetraacetic acid (EDTA) disodium salt 2-hydrate PanReac AppliChem Cat# 131669.1210 Sodium phosphate monobasic Sigma-Aldrich Cat# 71496-1 KG Sodium phosphate dibasic (for protein purification) Sigma-Aldrich Cat# 71640-1 KG Sodium phosphate dibasic dihydrate (for M9 preparation) Sigma-Aldrich Cat# 71643 Tris(2-carboxyethyl)phosphine Hydrochloride (TCEP) Apollo Scientific Cat# BIT0122 Imidazole PanReac AppliChem Cat# A1073,0500 HEPES PanReac AppliChem Cat# A1069,0500 Sodium hydroxide PanReac AppliChem Cat# 141929.1211 Potassium phosphate dibasic Sigma-Aldrich Cat# P3786-1 KG Potassium dihydrogen phosphate ACEF Cat# 001191-1 Bovine serum albumin Sigma-Aldrich Cat# A6003-25G Sodium sulfide Sigma-Aldrich Cat# 407410-10G Hydrochloric acid 37% PanReac AppliChem Cat# 131020.1212 Acetic acid glacial VWR BDH Chemicals Cat# 20104.334 Ninhydrin PanReac AppliChem Cat# A0902,0100 Ethanol 96% Vener – Sodium thiosulfate anhydrous (for enzymatic assays) Fluka Cat# 72049 Sodium thiosulfate pentahydrate (for media supplementation) Sigma-Aldrich Cat# 217247 O-acetyl-L-serine hydrochloride Sigma-Aldrich Cat# A6262-5G O-phospho-L-serine Sigma-Aldrich Cat# P0878-5G L-cysteine (for enzymatic assays) Sigma Cat# C-7755 L-cysteine hydrochloride monohydrate (for media supplementation) Fluka Sigma-Aldrich Cat# 30130 (Continued on next page) e1 iScience 29, 114304, January 16, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER L-serine Sigma-Aldrich Cat# 84959-25G Luria-Bertani (LB) broth Liofilchem S.r.l Cat# 610084 Agar (for protein expression) PanReac AppliChem Cat# A3477,0500 Agar (for solid M9 preparation) Liofilchem S.r.l Cat# 611001 Succinic acid disodium salt Sigma-Aldrich Cat# 224731 Magnesium sulfate heptahydrate Merck Cat# 1058860500 Magnesium chloride anhydrous BDH Limited Poole England Cat# 26123 Ammonium chloride Merck Cat# 1011451000 Calcium chloride dihydrate Sigma-Aldrich Cat# C3881-500G; Lot# 018K0680 Ampicillin sodium salt Sigma-Aldrich Cat# A9518; Lot# BCBZ9179 Nalidixic acid Sigma-Aldrich Cat# N8878-5G; Lot# 100K0122 Chloramphenicol Sigma-Aldrich Cat# C0378-25G; Lot# 0000127603 Tetracycline hydrochloride Sigma-Aldrich Cat# T3383-25G; Lot# 088K0680 Kanamycin sulfate Sigma-Aldrich Cat# K4000-25G; Lot# SLBR6873V Carbenicillin disodium salt Sigma-Aldrich Cat# C1389-1G; Lot# 0000097755 PaCysK This paper – PaCysM This paper – Precision Plus Protein TM Unstained Standards BIO-RAD Cat# 1610363 Tobacco Etch Virus (TEV) Protease This paper – FastDigest restriction enzyme_XbaI Thermo Fisher Scientific Cat# FD0684 FastDigest restriction enzyme_BamHI Thermo Fisher Scientific Cat# FD0054 FastDigest restriction enzyme_HindIII Thermo Fisher Scientific Cat# FD0504 FastDigest restriction enzyme_KpnI Thermo Fisher Scientific Cat# FD0524 FastDigest restriction enzyme_XhoI Thermo Fisher Scientific Cat# FD0694 Phusion TM High-Fidelity DNA Polymerase (2 u/μL) Thermo Fisher Scientific Cat# F530S T4 DNA Ligase, 100u Promega Italia Srl Cat# M1801 GoTaq(R) G2 DNA Polymerase, 100u Promega Italia Srl Cat# M7841 Critical commercial assays Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 PureYield(TM) Plasmid Miniprep System, 100 preps Promega Italia Srl Cat# A1223 H 2 S/SULF calibration kit Unisense A/S Cat# CALKIT-H2S; Lot# 1.06 Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 Deposited data SMP180 dataset Abdul-Gader et al. 96 https://doi.org/10.1093/bioinformatics/ btr234 Pseudomonas Genome Database SciCrunch Registry RRID:SCR_006590, http://www. pseudomonas.com/ Oligonucleotides See Table S3 for Oligonucleotides Recombinant DNA pET28a-TEV-PaCysK2709 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com pET28a-TEV-PaCysK0932 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com (Continued on next page) iScience 29, 114304, January 16, 2026 e2

    Expressing:

    Article Title: INF2 formin variants linked to human inherited kidney disease reprogram the transcriptome, causing mitotic chaos and cell death
    Article Snippet: .. Subsequently, the temperature was lowered to 20 °C, and recombinant protein expression was induced by adding 0.5 mM IPTG (Apollo Scientific, # BIMB1008) for 16 h. Cell lysates from HEK293T cells expressing GFP alone or GFP-centrins 1–3 or GFP-CaM were incubated with assay buffer (20 mM Tris–HCl, pH 7.8, 150 mM NaCl, 1% Nonidet P-40, 1% glycerol, 1 mM sodium orthovanadate, 0.1 mM PMSF, and 5 mM CaCl 2 ), supplemented with a commercial cocktail of protease inhibitors (Merck, # 11697498001) at 4 °C, alongside 30 μg of the indicated GST-fused proteins immobilized on glutathione-Sepharose 4B beads (Merck, # GE17-0756-01). ..

    Article Title: INF2 formin variants linked to human inherited kidney disease reprogram the transcriptome, causing mitotic chaos and cell death.
    Article Snippet: .. Subsequently, the temperature was lowered to 20 °C, and recombinant protein expression was induced by adding 0.5 mM IPTG (Apollo Scientific, # BIMB1008) for 16 h. Cell lysates from HEK293T cells expressing GFP alone or GFP-centrins 1–3 or GFP-CaM were incubated with assay buffer (20 mM Tris–HCl, pH 7.8, 150 mM NaCl, 1% Nonidet P-40, 1% glycerol, 1 mM sodium orthovanadate, 0.1 mM PMSF, and 5 mM CaCl2), supplemented with a commercial cocktail of protease inhibitors (Merck, # 11697498001) at 4 °C, alongside 30 μg of the indicated GST-fused proteins immobilized on glutathioneSepharose 4B beads (Merck, # GE17-0756-01). ..

    Article Title: De novo cysteine biosynthesis in Pseudomonas aeruginosa: characterization of the two main cysteine synthase isoforms
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains See Table 3 for bacterial strains Chemicals, peptides, and recombinant proteins Yeast extract PanReac AppliChem Cat# A1552,1000 Tryptone PanReac AppliChem Cat# A1553,1000 Sodium chloride (for protein expression and purification) PanReac AppliChem Cat# 131659.1211 Sodium chloride (for M9 preparation) Sigma-Aldrich Cat# 746398 Phosphate buffered saline Sigma-Aldrich Cat# P4417-50TAB D-(+)-Glucose Sigma Life Science Cat# G8270-10 KG Isopropil β-D-1-tiogalattopiranoside, IPTG, Isopropil β-D-tiogalattoside (IPTG) Apollo Scientific Cat# BIMB1008 Phenylmethanesulphonyl fluoride (PMSF) Apollo Scientific Cat# PC6222M Benzamidine Fluka Cat# 12072 Pepstatin A PanReac AppliChem Cat# A2205,0010 Pyridoxal-5 ′ -phosphate hydrate (PLP) Sigma-Aldrich Cat# P9255-5G Lysozime from chicken egg white Sigma-Aldrich Cat# 62971-10G-F Ethylenediaminetetraacetic acid (EDTA) disodium salt 2-hydrate PanReac AppliChem Cat# 131669.1210 Sodium phosphate monobasic Sigma-Aldrich Cat# 71496-1 KG Sodium phosphate dibasic (for protein purification) Sigma-Aldrich Cat# 71640-1 KG Sodium phosphate dibasic dihydrate (for M9 preparation) Sigma-Aldrich Cat# 71643 Tris(2-carboxyethyl)phosphine Hydrochloride (TCEP) Apollo Scientific Cat# BIT0122 Imidazole PanReac AppliChem Cat# A1073,0500 HEPES PanReac AppliChem Cat# A1069,0500 Sodium hydroxide PanReac AppliChem Cat# 141929.1211 Potassium phosphate dibasic Sigma-Aldrich Cat# P3786-1 KG Potassium dihydrogen phosphate ACEF Cat# 001191-1 Bovine serum albumin Sigma-Aldrich Cat# A6003-25G Sodium sulfide Sigma-Aldrich Cat# 407410-10G Hydrochloric acid 37% PanReac AppliChem Cat# 131020.1212 Acetic acid glacial VWR BDH Chemicals Cat# 20104.334 Ninhydrin PanReac AppliChem Cat# A0902,0100 Ethanol 96% Vener – Sodium thiosulfate anhydrous (for enzymatic assays) Fluka Cat# 72049 Sodium thiosulfate pentahydrate (for media supplementation) Sigma-Aldrich Cat# 217247 O-acetyl-L-serine hydrochloride Sigma-Aldrich Cat# A6262-5G O-phospho-L-serine Sigma-Aldrich Cat# P0878-5G L-cysteine (for enzymatic assays) Sigma Cat# C-7755 L-cysteine hydrochloride monohydrate (for media supplementation) Fluka Sigma-Aldrich Cat# 30130 (Continued on next page) e1 iScience 29, 114304, January 16, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER L-serine Sigma-Aldrich Cat# 84959-25G Luria-Bertani (LB) broth Liofilchem S.r.l Cat# 610084 Agar (for protein expression) PanReac AppliChem Cat# A3477,0500 Agar (for solid M9 preparation) Liofilchem S.r.l Cat# 611001 Succinic acid disodium salt Sigma-Aldrich Cat# 224731 Magnesium sulfate heptahydrate Merck Cat# 1058860500 Magnesium chloride anhydrous BDH Limited Poole England Cat# 26123 Ammonium chloride Merck Cat# 1011451000 Calcium chloride dihydrate Sigma-Aldrich Cat# C3881-500G; Lot# 018K0680 Ampicillin sodium salt Sigma-Aldrich Cat# A9518; Lot# BCBZ9179 Nalidixic acid Sigma-Aldrich Cat# N8878-5G; Lot# 100K0122 Chloramphenicol Sigma-Aldrich Cat# C0378-25G; Lot# 0000127603 Tetracycline hydrochloride Sigma-Aldrich Cat# T3383-25G; Lot# 088K0680 Kanamycin sulfate Sigma-Aldrich Cat# K4000-25G; Lot# SLBR6873V Carbenicillin disodium salt Sigma-Aldrich Cat# C1389-1G; Lot# 0000097755 PaCysK This paper – PaCysM This paper – Precision Plus Protein TM Unstained Standards BIO-RAD Cat# 1610363 Tobacco Etch Virus (TEV) Protease This paper – FastDigest restriction enzyme_XbaI Thermo Fisher Scientific Cat# FD0684 FastDigest restriction enzyme_BamHI Thermo Fisher Scientific Cat# FD0054 FastDigest restriction enzyme_HindIII Thermo Fisher Scientific Cat# FD0504 FastDigest restriction enzyme_KpnI Thermo Fisher Scientific Cat# FD0524 FastDigest restriction enzyme_XhoI Thermo Fisher Scientific Cat# FD0694 Phusion TM High-Fidelity DNA Polymerase (2 u/μL) Thermo Fisher Scientific Cat# F530S T4 DNA Ligase, 100u Promega Italia Srl Cat# M1801 GoTaq(R) G2 DNA Polymerase, 100u Promega Italia Srl Cat# M7841 Critical commercial assays Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 PureYield(TM) Plasmid Miniprep System, 100 preps Promega Italia Srl Cat# A1223 H 2 S/SULF calibration kit Unisense A/S Cat# CALKIT-H2S; Lot# 1.06 Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 Deposited data SMP180 dataset Abdul-Gader et al. 96 https://doi.org/10.1093/bioinformatics/ btr234 Pseudomonas Genome Database SciCrunch Registry RRID:SCR_006590, http://www. pseudomonas.com/ Oligonucleotides See Table S3 for Oligonucleotides Recombinant DNA pET28a-TEV-PaCysK2709 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com pET28a-TEV-PaCysK0932 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com (Continued on next page) iScience 29, 114304, January 16, 2026 e2

    Incubation:

    Article Title: INF2 formin variants linked to human inherited kidney disease reprogram the transcriptome, causing mitotic chaos and cell death
    Article Snippet: .. Subsequently, the temperature was lowered to 20 °C, and recombinant protein expression was induced by adding 0.5 mM IPTG (Apollo Scientific, # BIMB1008) for 16 h. Cell lysates from HEK293T cells expressing GFP alone or GFP-centrins 1–3 or GFP-CaM were incubated with assay buffer (20 mM Tris–HCl, pH 7.8, 150 mM NaCl, 1% Nonidet P-40, 1% glycerol, 1 mM sodium orthovanadate, 0.1 mM PMSF, and 5 mM CaCl 2 ), supplemented with a commercial cocktail of protease inhibitors (Merck, # 11697498001) at 4 °C, alongside 30 μg of the indicated GST-fused proteins immobilized on glutathione-Sepharose 4B beads (Merck, # GE17-0756-01). ..

    Article Title: INF2 formin variants linked to human inherited kidney disease reprogram the transcriptome, causing mitotic chaos and cell death.
    Article Snippet: .. Subsequently, the temperature was lowered to 20 °C, and recombinant protein expression was induced by adding 0.5 mM IPTG (Apollo Scientific, # BIMB1008) for 16 h. Cell lysates from HEK293T cells expressing GFP alone or GFP-centrins 1–3 or GFP-CaM were incubated with assay buffer (20 mM Tris–HCl, pH 7.8, 150 mM NaCl, 1% Nonidet P-40, 1% glycerol, 1 mM sodium orthovanadate, 0.1 mM PMSF, and 5 mM CaCl2), supplemented with a commercial cocktail of protease inhibitors (Merck, # 11697498001) at 4 °C, alongside 30 μg of the indicated GST-fused proteins immobilized on glutathioneSepharose 4B beads (Merck, # GE17-0756-01). ..

    other:

    Article Title: Cell constriction requires processive septal peptidoglycan synthase movement independent of FtsZ treadmilling in Staphylococcus aureus
    Article Snippet: Unless otherwise specified, 5-bromo-4-chloro-3-indolyl β- d -galactopyranoside (X-Gal, Apollo Scientific) was used at 100 μg ml −1 , IPTG (Apollo Scientific) was used at 0.5 mM and Atc (Sigma-Aldrich) was used at 2 ng ml −1 .

    Virus:

    Article Title: De novo cysteine biosynthesis in Pseudomonas aeruginosa: characterization of the two main cysteine synthase isoforms
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains See Table 3 for bacterial strains Chemicals, peptides, and recombinant proteins Yeast extract PanReac AppliChem Cat# A1552,1000 Tryptone PanReac AppliChem Cat# A1553,1000 Sodium chloride (for protein expression and purification) PanReac AppliChem Cat# 131659.1211 Sodium chloride (for M9 preparation) Sigma-Aldrich Cat# 746398 Phosphate buffered saline Sigma-Aldrich Cat# P4417-50TAB D-(+)-Glucose Sigma Life Science Cat# G8270-10 KG Isopropil β-D-1-tiogalattopiranoside, IPTG, Isopropil β-D-tiogalattoside (IPTG) Apollo Scientific Cat# BIMB1008 Phenylmethanesulphonyl fluoride (PMSF) Apollo Scientific Cat# PC6222M Benzamidine Fluka Cat# 12072 Pepstatin A PanReac AppliChem Cat# A2205,0010 Pyridoxal-5 ′ -phosphate hydrate (PLP) Sigma-Aldrich Cat# P9255-5G Lysozime from chicken egg white Sigma-Aldrich Cat# 62971-10G-F Ethylenediaminetetraacetic acid (EDTA) disodium salt 2-hydrate PanReac AppliChem Cat# 131669.1210 Sodium phosphate monobasic Sigma-Aldrich Cat# 71496-1 KG Sodium phosphate dibasic (for protein purification) Sigma-Aldrich Cat# 71640-1 KG Sodium phosphate dibasic dihydrate (for M9 preparation) Sigma-Aldrich Cat# 71643 Tris(2-carboxyethyl)phosphine Hydrochloride (TCEP) Apollo Scientific Cat# BIT0122 Imidazole PanReac AppliChem Cat# A1073,0500 HEPES PanReac AppliChem Cat# A1069,0500 Sodium hydroxide PanReac AppliChem Cat# 141929.1211 Potassium phosphate dibasic Sigma-Aldrich Cat# P3786-1 KG Potassium dihydrogen phosphate ACEF Cat# 001191-1 Bovine serum albumin Sigma-Aldrich Cat# A6003-25G Sodium sulfide Sigma-Aldrich Cat# 407410-10G Hydrochloric acid 37% PanReac AppliChem Cat# 131020.1212 Acetic acid glacial VWR BDH Chemicals Cat# 20104.334 Ninhydrin PanReac AppliChem Cat# A0902,0100 Ethanol 96% Vener – Sodium thiosulfate anhydrous (for enzymatic assays) Fluka Cat# 72049 Sodium thiosulfate pentahydrate (for media supplementation) Sigma-Aldrich Cat# 217247 O-acetyl-L-serine hydrochloride Sigma-Aldrich Cat# A6262-5G O-phospho-L-serine Sigma-Aldrich Cat# P0878-5G L-cysteine (for enzymatic assays) Sigma Cat# C-7755 L-cysteine hydrochloride monohydrate (for media supplementation) Fluka Sigma-Aldrich Cat# 30130 (Continued on next page) e1 iScience 29, 114304, January 16, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER L-serine Sigma-Aldrich Cat# 84959-25G Luria-Bertani (LB) broth Liofilchem S.r.l Cat# 610084 Agar (for protein expression) PanReac AppliChem Cat# A3477,0500 Agar (for solid M9 preparation) Liofilchem S.r.l Cat# 611001 Succinic acid disodium salt Sigma-Aldrich Cat# 224731 Magnesium sulfate heptahydrate Merck Cat# 1058860500 Magnesium chloride anhydrous BDH Limited Poole England Cat# 26123 Ammonium chloride Merck Cat# 1011451000 Calcium chloride dihydrate Sigma-Aldrich Cat# C3881-500G; Lot# 018K0680 Ampicillin sodium salt Sigma-Aldrich Cat# A9518; Lot# BCBZ9179 Nalidixic acid Sigma-Aldrich Cat# N8878-5G; Lot# 100K0122 Chloramphenicol Sigma-Aldrich Cat# C0378-25G; Lot# 0000127603 Tetracycline hydrochloride Sigma-Aldrich Cat# T3383-25G; Lot# 088K0680 Kanamycin sulfate Sigma-Aldrich Cat# K4000-25G; Lot# SLBR6873V Carbenicillin disodium salt Sigma-Aldrich Cat# C1389-1G; Lot# 0000097755 PaCysK This paper – PaCysM This paper – Precision Plus Protein TM Unstained Standards BIO-RAD Cat# 1610363 Tobacco Etch Virus (TEV) Protease This paper – FastDigest restriction enzyme_XbaI Thermo Fisher Scientific Cat# FD0684 FastDigest restriction enzyme_BamHI Thermo Fisher Scientific Cat# FD0054 FastDigest restriction enzyme_HindIII Thermo Fisher Scientific Cat# FD0504 FastDigest restriction enzyme_KpnI Thermo Fisher Scientific Cat# FD0524 FastDigest restriction enzyme_XhoI Thermo Fisher Scientific Cat# FD0694 Phusion TM High-Fidelity DNA Polymerase (2 u/μL) Thermo Fisher Scientific Cat# F530S T4 DNA Ligase, 100u Promega Italia Srl Cat# M1801 GoTaq(R) G2 DNA Polymerase, 100u Promega Italia Srl Cat# M7841 Critical commercial assays Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 PureYield(TM) Plasmid Miniprep System, 100 preps Promega Italia Srl Cat# A1223 H 2 S/SULF calibration kit Unisense A/S Cat# CALKIT-H2S; Lot# 1.06 Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 Deposited data SMP180 dataset Abdul-Gader et al. 96 https://doi.org/10.1093/bioinformatics/ btr234 Pseudomonas Genome Database SciCrunch Registry RRID:SCR_006590, http://www. pseudomonas.com/ Oligonucleotides See Table S3 for Oligonucleotides Recombinant DNA pET28a-TEV-PaCysK2709 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com pET28a-TEV-PaCysK0932 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com (Continued on next page) iScience 29, 114304, January 16, 2026 e2

    Purification:

    Article Title: De novo cysteine biosynthesis in Pseudomonas aeruginosa: characterization of the two main cysteine synthase isoforms
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains See Table 3 for bacterial strains Chemicals, peptides, and recombinant proteins Yeast extract PanReac AppliChem Cat# A1552,1000 Tryptone PanReac AppliChem Cat# A1553,1000 Sodium chloride (for protein expression and purification) PanReac AppliChem Cat# 131659.1211 Sodium chloride (for M9 preparation) Sigma-Aldrich Cat# 746398 Phosphate buffered saline Sigma-Aldrich Cat# P4417-50TAB D-(+)-Glucose Sigma Life Science Cat# G8270-10 KG Isopropil β-D-1-tiogalattopiranoside, IPTG, Isopropil β-D-tiogalattoside (IPTG) Apollo Scientific Cat# BIMB1008 Phenylmethanesulphonyl fluoride (PMSF) Apollo Scientific Cat# PC6222M Benzamidine Fluka Cat# 12072 Pepstatin A PanReac AppliChem Cat# A2205,0010 Pyridoxal-5 ′ -phosphate hydrate (PLP) Sigma-Aldrich Cat# P9255-5G Lysozime from chicken egg white Sigma-Aldrich Cat# 62971-10G-F Ethylenediaminetetraacetic acid (EDTA) disodium salt 2-hydrate PanReac AppliChem Cat# 131669.1210 Sodium phosphate monobasic Sigma-Aldrich Cat# 71496-1 KG Sodium phosphate dibasic (for protein purification) Sigma-Aldrich Cat# 71640-1 KG Sodium phosphate dibasic dihydrate (for M9 preparation) Sigma-Aldrich Cat# 71643 Tris(2-carboxyethyl)phosphine Hydrochloride (TCEP) Apollo Scientific Cat# BIT0122 Imidazole PanReac AppliChem Cat# A1073,0500 HEPES PanReac AppliChem Cat# A1069,0500 Sodium hydroxide PanReac AppliChem Cat# 141929.1211 Potassium phosphate dibasic Sigma-Aldrich Cat# P3786-1 KG Potassium dihydrogen phosphate ACEF Cat# 001191-1 Bovine serum albumin Sigma-Aldrich Cat# A6003-25G Sodium sulfide Sigma-Aldrich Cat# 407410-10G Hydrochloric acid 37% PanReac AppliChem Cat# 131020.1212 Acetic acid glacial VWR BDH Chemicals Cat# 20104.334 Ninhydrin PanReac AppliChem Cat# A0902,0100 Ethanol 96% Vener – Sodium thiosulfate anhydrous (for enzymatic assays) Fluka Cat# 72049 Sodium thiosulfate pentahydrate (for media supplementation) Sigma-Aldrich Cat# 217247 O-acetyl-L-serine hydrochloride Sigma-Aldrich Cat# A6262-5G O-phospho-L-serine Sigma-Aldrich Cat# P0878-5G L-cysteine (for enzymatic assays) Sigma Cat# C-7755 L-cysteine hydrochloride monohydrate (for media supplementation) Fluka Sigma-Aldrich Cat# 30130 (Continued on next page) e1 iScience 29, 114304, January 16, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER L-serine Sigma-Aldrich Cat# 84959-25G Luria-Bertani (LB) broth Liofilchem S.r.l Cat# 610084 Agar (for protein expression) PanReac AppliChem Cat# A3477,0500 Agar (for solid M9 preparation) Liofilchem S.r.l Cat# 611001 Succinic acid disodium salt Sigma-Aldrich Cat# 224731 Magnesium sulfate heptahydrate Merck Cat# 1058860500 Magnesium chloride anhydrous BDH Limited Poole England Cat# 26123 Ammonium chloride Merck Cat# 1011451000 Calcium chloride dihydrate Sigma-Aldrich Cat# C3881-500G; Lot# 018K0680 Ampicillin sodium salt Sigma-Aldrich Cat# A9518; Lot# BCBZ9179 Nalidixic acid Sigma-Aldrich Cat# N8878-5G; Lot# 100K0122 Chloramphenicol Sigma-Aldrich Cat# C0378-25G; Lot# 0000127603 Tetracycline hydrochloride Sigma-Aldrich Cat# T3383-25G; Lot# 088K0680 Kanamycin sulfate Sigma-Aldrich Cat# K4000-25G; Lot# SLBR6873V Carbenicillin disodium salt Sigma-Aldrich Cat# C1389-1G; Lot# 0000097755 PaCysK This paper – PaCysM This paper – Precision Plus Protein TM Unstained Standards BIO-RAD Cat# 1610363 Tobacco Etch Virus (TEV) Protease This paper – FastDigest restriction enzyme_XbaI Thermo Fisher Scientific Cat# FD0684 FastDigest restriction enzyme_BamHI Thermo Fisher Scientific Cat# FD0054 FastDigest restriction enzyme_HindIII Thermo Fisher Scientific Cat# FD0504 FastDigest restriction enzyme_KpnI Thermo Fisher Scientific Cat# FD0524 FastDigest restriction enzyme_XhoI Thermo Fisher Scientific Cat# FD0694 Phusion TM High-Fidelity DNA Polymerase (2 u/μL) Thermo Fisher Scientific Cat# F530S T4 DNA Ligase, 100u Promega Italia Srl Cat# M1801 GoTaq(R) G2 DNA Polymerase, 100u Promega Italia Srl Cat# M7841 Critical commercial assays Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 PureYield(TM) Plasmid Miniprep System, 100 preps Promega Italia Srl Cat# A1223 H 2 S/SULF calibration kit Unisense A/S Cat# CALKIT-H2S; Lot# 1.06 Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 Deposited data SMP180 dataset Abdul-Gader et al. 96 https://doi.org/10.1093/bioinformatics/ btr234 Pseudomonas Genome Database SciCrunch Registry RRID:SCR_006590, http://www. pseudomonas.com/ Oligonucleotides See Table S3 for Oligonucleotides Recombinant DNA pET28a-TEV-PaCysK2709 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com pET28a-TEV-PaCysK0932 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com (Continued on next page) iScience 29, 114304, January 16, 2026 e2

    Saline:

    Article Title: De novo cysteine biosynthesis in Pseudomonas aeruginosa: characterization of the two main cysteine synthase isoforms
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains See Table 3 for bacterial strains Chemicals, peptides, and recombinant proteins Yeast extract PanReac AppliChem Cat# A1552,1000 Tryptone PanReac AppliChem Cat# A1553,1000 Sodium chloride (for protein expression and purification) PanReac AppliChem Cat# 131659.1211 Sodium chloride (for M9 preparation) Sigma-Aldrich Cat# 746398 Phosphate buffered saline Sigma-Aldrich Cat# P4417-50TAB D-(+)-Glucose Sigma Life Science Cat# G8270-10 KG Isopropil β-D-1-tiogalattopiranoside, IPTG, Isopropil β-D-tiogalattoside (IPTG) Apollo Scientific Cat# BIMB1008 Phenylmethanesulphonyl fluoride (PMSF) Apollo Scientific Cat# PC6222M Benzamidine Fluka Cat# 12072 Pepstatin A PanReac AppliChem Cat# A2205,0010 Pyridoxal-5 ′ -phosphate hydrate (PLP) Sigma-Aldrich Cat# P9255-5G Lysozime from chicken egg white Sigma-Aldrich Cat# 62971-10G-F Ethylenediaminetetraacetic acid (EDTA) disodium salt 2-hydrate PanReac AppliChem Cat# 131669.1210 Sodium phosphate monobasic Sigma-Aldrich Cat# 71496-1 KG Sodium phosphate dibasic (for protein purification) Sigma-Aldrich Cat# 71640-1 KG Sodium phosphate dibasic dihydrate (for M9 preparation) Sigma-Aldrich Cat# 71643 Tris(2-carboxyethyl)phosphine Hydrochloride (TCEP) Apollo Scientific Cat# BIT0122 Imidazole PanReac AppliChem Cat# A1073,0500 HEPES PanReac AppliChem Cat# A1069,0500 Sodium hydroxide PanReac AppliChem Cat# 141929.1211 Potassium phosphate dibasic Sigma-Aldrich Cat# P3786-1 KG Potassium dihydrogen phosphate ACEF Cat# 001191-1 Bovine serum albumin Sigma-Aldrich Cat# A6003-25G Sodium sulfide Sigma-Aldrich Cat# 407410-10G Hydrochloric acid 37% PanReac AppliChem Cat# 131020.1212 Acetic acid glacial VWR BDH Chemicals Cat# 20104.334 Ninhydrin PanReac AppliChem Cat# A0902,0100 Ethanol 96% Vener – Sodium thiosulfate anhydrous (for enzymatic assays) Fluka Cat# 72049 Sodium thiosulfate pentahydrate (for media supplementation) Sigma-Aldrich Cat# 217247 O-acetyl-L-serine hydrochloride Sigma-Aldrich Cat# A6262-5G O-phospho-L-serine Sigma-Aldrich Cat# P0878-5G L-cysteine (for enzymatic assays) Sigma Cat# C-7755 L-cysteine hydrochloride monohydrate (for media supplementation) Fluka Sigma-Aldrich Cat# 30130 (Continued on next page) e1 iScience 29, 114304, January 16, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER L-serine Sigma-Aldrich Cat# 84959-25G Luria-Bertani (LB) broth Liofilchem S.r.l Cat# 610084 Agar (for protein expression) PanReac AppliChem Cat# A3477,0500 Agar (for solid M9 preparation) Liofilchem S.r.l Cat# 611001 Succinic acid disodium salt Sigma-Aldrich Cat# 224731 Magnesium sulfate heptahydrate Merck Cat# 1058860500 Magnesium chloride anhydrous BDH Limited Poole England Cat# 26123 Ammonium chloride Merck Cat# 1011451000 Calcium chloride dihydrate Sigma-Aldrich Cat# C3881-500G; Lot# 018K0680 Ampicillin sodium salt Sigma-Aldrich Cat# A9518; Lot# BCBZ9179 Nalidixic acid Sigma-Aldrich Cat# N8878-5G; Lot# 100K0122 Chloramphenicol Sigma-Aldrich Cat# C0378-25G; Lot# 0000127603 Tetracycline hydrochloride Sigma-Aldrich Cat# T3383-25G; Lot# 088K0680 Kanamycin sulfate Sigma-Aldrich Cat# K4000-25G; Lot# SLBR6873V Carbenicillin disodium salt Sigma-Aldrich Cat# C1389-1G; Lot# 0000097755 PaCysK This paper – PaCysM This paper – Precision Plus Protein TM Unstained Standards BIO-RAD Cat# 1610363 Tobacco Etch Virus (TEV) Protease This paper – FastDigest restriction enzyme_XbaI Thermo Fisher Scientific Cat# FD0684 FastDigest restriction enzyme_BamHI Thermo Fisher Scientific Cat# FD0054 FastDigest restriction enzyme_HindIII Thermo Fisher Scientific Cat# FD0504 FastDigest restriction enzyme_KpnI Thermo Fisher Scientific Cat# FD0524 FastDigest restriction enzyme_XhoI Thermo Fisher Scientific Cat# FD0694 Phusion TM High-Fidelity DNA Polymerase (2 u/μL) Thermo Fisher Scientific Cat# F530S T4 DNA Ligase, 100u Promega Italia Srl Cat# M1801 GoTaq(R) G2 DNA Polymerase, 100u Promega Italia Srl Cat# M7841 Critical commercial assays Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 PureYield(TM) Plasmid Miniprep System, 100 preps Promega Italia Srl Cat# A1223 H 2 S/SULF calibration kit Unisense A/S Cat# CALKIT-H2S; Lot# 1.06 Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 Deposited data SMP180 dataset Abdul-Gader et al. 96 https://doi.org/10.1093/bioinformatics/ btr234 Pseudomonas Genome Database SciCrunch Registry RRID:SCR_006590, http://www. pseudomonas.com/ Oligonucleotides See Table S3 for Oligonucleotides Recombinant DNA pET28a-TEV-PaCysK2709 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com pET28a-TEV-PaCysK0932 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com (Continued on next page) iScience 29, 114304, January 16, 2026 e2

    Plasmid Purification:

    Article Title: De novo cysteine biosynthesis in Pseudomonas aeruginosa: characterization of the two main cysteine synthase isoforms
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains See Table 3 for bacterial strains Chemicals, peptides, and recombinant proteins Yeast extract PanReac AppliChem Cat# A1552,1000 Tryptone PanReac AppliChem Cat# A1553,1000 Sodium chloride (for protein expression and purification) PanReac AppliChem Cat# 131659.1211 Sodium chloride (for M9 preparation) Sigma-Aldrich Cat# 746398 Phosphate buffered saline Sigma-Aldrich Cat# P4417-50TAB D-(+)-Glucose Sigma Life Science Cat# G8270-10 KG Isopropil β-D-1-tiogalattopiranoside, IPTG, Isopropil β-D-tiogalattoside (IPTG) Apollo Scientific Cat# BIMB1008 Phenylmethanesulphonyl fluoride (PMSF) Apollo Scientific Cat# PC6222M Benzamidine Fluka Cat# 12072 Pepstatin A PanReac AppliChem Cat# A2205,0010 Pyridoxal-5 ′ -phosphate hydrate (PLP) Sigma-Aldrich Cat# P9255-5G Lysozime from chicken egg white Sigma-Aldrich Cat# 62971-10G-F Ethylenediaminetetraacetic acid (EDTA) disodium salt 2-hydrate PanReac AppliChem Cat# 131669.1210 Sodium phosphate monobasic Sigma-Aldrich Cat# 71496-1 KG Sodium phosphate dibasic (for protein purification) Sigma-Aldrich Cat# 71640-1 KG Sodium phosphate dibasic dihydrate (for M9 preparation) Sigma-Aldrich Cat# 71643 Tris(2-carboxyethyl)phosphine Hydrochloride (TCEP) Apollo Scientific Cat# BIT0122 Imidazole PanReac AppliChem Cat# A1073,0500 HEPES PanReac AppliChem Cat# A1069,0500 Sodium hydroxide PanReac AppliChem Cat# 141929.1211 Potassium phosphate dibasic Sigma-Aldrich Cat# P3786-1 KG Potassium dihydrogen phosphate ACEF Cat# 001191-1 Bovine serum albumin Sigma-Aldrich Cat# A6003-25G Sodium sulfide Sigma-Aldrich Cat# 407410-10G Hydrochloric acid 37% PanReac AppliChem Cat# 131020.1212 Acetic acid glacial VWR BDH Chemicals Cat# 20104.334 Ninhydrin PanReac AppliChem Cat# A0902,0100 Ethanol 96% Vener – Sodium thiosulfate anhydrous (for enzymatic assays) Fluka Cat# 72049 Sodium thiosulfate pentahydrate (for media supplementation) Sigma-Aldrich Cat# 217247 O-acetyl-L-serine hydrochloride Sigma-Aldrich Cat# A6262-5G O-phospho-L-serine Sigma-Aldrich Cat# P0878-5G L-cysteine (for enzymatic assays) Sigma Cat# C-7755 L-cysteine hydrochloride monohydrate (for media supplementation) Fluka Sigma-Aldrich Cat# 30130 (Continued on next page) e1 iScience 29, 114304, January 16, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER L-serine Sigma-Aldrich Cat# 84959-25G Luria-Bertani (LB) broth Liofilchem S.r.l Cat# 610084 Agar (for protein expression) PanReac AppliChem Cat# A3477,0500 Agar (for solid M9 preparation) Liofilchem S.r.l Cat# 611001 Succinic acid disodium salt Sigma-Aldrich Cat# 224731 Magnesium sulfate heptahydrate Merck Cat# 1058860500 Magnesium chloride anhydrous BDH Limited Poole England Cat# 26123 Ammonium chloride Merck Cat# 1011451000 Calcium chloride dihydrate Sigma-Aldrich Cat# C3881-500G; Lot# 018K0680 Ampicillin sodium salt Sigma-Aldrich Cat# A9518; Lot# BCBZ9179 Nalidixic acid Sigma-Aldrich Cat# N8878-5G; Lot# 100K0122 Chloramphenicol Sigma-Aldrich Cat# C0378-25G; Lot# 0000127603 Tetracycline hydrochloride Sigma-Aldrich Cat# T3383-25G; Lot# 088K0680 Kanamycin sulfate Sigma-Aldrich Cat# K4000-25G; Lot# SLBR6873V Carbenicillin disodium salt Sigma-Aldrich Cat# C1389-1G; Lot# 0000097755 PaCysK This paper – PaCysM This paper – Precision Plus Protein TM Unstained Standards BIO-RAD Cat# 1610363 Tobacco Etch Virus (TEV) Protease This paper – FastDigest restriction enzyme_XbaI Thermo Fisher Scientific Cat# FD0684 FastDigest restriction enzyme_BamHI Thermo Fisher Scientific Cat# FD0054 FastDigest restriction enzyme_HindIII Thermo Fisher Scientific Cat# FD0504 FastDigest restriction enzyme_KpnI Thermo Fisher Scientific Cat# FD0524 FastDigest restriction enzyme_XhoI Thermo Fisher Scientific Cat# FD0694 Phusion TM High-Fidelity DNA Polymerase (2 u/μL) Thermo Fisher Scientific Cat# F530S T4 DNA Ligase, 100u Promega Italia Srl Cat# M1801 GoTaq(R) G2 DNA Polymerase, 100u Promega Italia Srl Cat# M7841 Critical commercial assays Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 PureYield(TM) Plasmid Miniprep System, 100 preps Promega Italia Srl Cat# A1223 H 2 S/SULF calibration kit Unisense A/S Cat# CALKIT-H2S; Lot# 1.06 Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 Deposited data SMP180 dataset Abdul-Gader et al. 96 https://doi.org/10.1093/bioinformatics/ btr234 Pseudomonas Genome Database SciCrunch Registry RRID:SCR_006590, http://www. pseudomonas.com/ Oligonucleotides See Table S3 for Oligonucleotides Recombinant DNA pET28a-TEV-PaCysK2709 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com pET28a-TEV-PaCysK0932 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com (Continued on next page) iScience 29, 114304, January 16, 2026 e2

    Protein Purification:

    Article Title: De novo cysteine biosynthesis in Pseudomonas aeruginosa: characterization of the two main cysteine synthase isoforms
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Bacterial and virus strains See Table 3 for bacterial strains Chemicals, peptides, and recombinant proteins Yeast extract PanReac AppliChem Cat# A1552,1000 Tryptone PanReac AppliChem Cat# A1553,1000 Sodium chloride (for protein expression and purification) PanReac AppliChem Cat# 131659.1211 Sodium chloride (for M9 preparation) Sigma-Aldrich Cat# 746398 Phosphate buffered saline Sigma-Aldrich Cat# P4417-50TAB D-(+)-Glucose Sigma Life Science Cat# G8270-10 KG Isopropil β-D-1-tiogalattopiranoside, IPTG, Isopropil β-D-tiogalattoside (IPTG) Apollo Scientific Cat# BIMB1008 Phenylmethanesulphonyl fluoride (PMSF) Apollo Scientific Cat# PC6222M Benzamidine Fluka Cat# 12072 Pepstatin A PanReac AppliChem Cat# A2205,0010 Pyridoxal-5 ′ -phosphate hydrate (PLP) Sigma-Aldrich Cat# P9255-5G Lysozime from chicken egg white Sigma-Aldrich Cat# 62971-10G-F Ethylenediaminetetraacetic acid (EDTA) disodium salt 2-hydrate PanReac AppliChem Cat# 131669.1210 Sodium phosphate monobasic Sigma-Aldrich Cat# 71496-1 KG Sodium phosphate dibasic (for protein purification) Sigma-Aldrich Cat# 71640-1 KG Sodium phosphate dibasic dihydrate (for M9 preparation) Sigma-Aldrich Cat# 71643 Tris(2-carboxyethyl)phosphine Hydrochloride (TCEP) Apollo Scientific Cat# BIT0122 Imidazole PanReac AppliChem Cat# A1073,0500 HEPES PanReac AppliChem Cat# A1069,0500 Sodium hydroxide PanReac AppliChem Cat# 141929.1211 Potassium phosphate dibasic Sigma-Aldrich Cat# P3786-1 KG Potassium dihydrogen phosphate ACEF Cat# 001191-1 Bovine serum albumin Sigma-Aldrich Cat# A6003-25G Sodium sulfide Sigma-Aldrich Cat# 407410-10G Hydrochloric acid 37% PanReac AppliChem Cat# 131020.1212 Acetic acid glacial VWR BDH Chemicals Cat# 20104.334 Ninhydrin PanReac AppliChem Cat# A0902,0100 Ethanol 96% Vener – Sodium thiosulfate anhydrous (for enzymatic assays) Fluka Cat# 72049 Sodium thiosulfate pentahydrate (for media supplementation) Sigma-Aldrich Cat# 217247 O-acetyl-L-serine hydrochloride Sigma-Aldrich Cat# A6262-5G O-phospho-L-serine Sigma-Aldrich Cat# P0878-5G L-cysteine (for enzymatic assays) Sigma Cat# C-7755 L-cysteine hydrochloride monohydrate (for media supplementation) Fluka Sigma-Aldrich Cat# 30130 (Continued on next page) e1 iScience 29, 114304, January 16, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER L-serine Sigma-Aldrich Cat# 84959-25G Luria-Bertani (LB) broth Liofilchem S.r.l Cat# 610084 Agar (for protein expression) PanReac AppliChem Cat# A3477,0500 Agar (for solid M9 preparation) Liofilchem S.r.l Cat# 611001 Succinic acid disodium salt Sigma-Aldrich Cat# 224731 Magnesium sulfate heptahydrate Merck Cat# 1058860500 Magnesium chloride anhydrous BDH Limited Poole England Cat# 26123 Ammonium chloride Merck Cat# 1011451000 Calcium chloride dihydrate Sigma-Aldrich Cat# C3881-500G; Lot# 018K0680 Ampicillin sodium salt Sigma-Aldrich Cat# A9518; Lot# BCBZ9179 Nalidixic acid Sigma-Aldrich Cat# N8878-5G; Lot# 100K0122 Chloramphenicol Sigma-Aldrich Cat# C0378-25G; Lot# 0000127603 Tetracycline hydrochloride Sigma-Aldrich Cat# T3383-25G; Lot# 088K0680 Kanamycin sulfate Sigma-Aldrich Cat# K4000-25G; Lot# SLBR6873V Carbenicillin disodium salt Sigma-Aldrich Cat# C1389-1G; Lot# 0000097755 PaCysK This paper – PaCysM This paper – Precision Plus Protein TM Unstained Standards BIO-RAD Cat# 1610363 Tobacco Etch Virus (TEV) Protease This paper – FastDigest restriction enzyme_XbaI Thermo Fisher Scientific Cat# FD0684 FastDigest restriction enzyme_BamHI Thermo Fisher Scientific Cat# FD0054 FastDigest restriction enzyme_HindIII Thermo Fisher Scientific Cat# FD0504 FastDigest restriction enzyme_KpnI Thermo Fisher Scientific Cat# FD0524 FastDigest restriction enzyme_XhoI Thermo Fisher Scientific Cat# FD0694 Phusion TM High-Fidelity DNA Polymerase (2 u/μL) Thermo Fisher Scientific Cat# F530S T4 DNA Ligase, 100u Promega Italia Srl Cat# M1801 GoTaq(R) G2 DNA Polymerase, 100u Promega Italia Srl Cat# M7841 Critical commercial assays Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 PureYield(TM) Plasmid Miniprep System, 100 preps Promega Italia Srl Cat# A1223 H 2 S/SULF calibration kit Unisense A/S Cat# CALKIT-H2S; Lot# 1.06 Wizard(R) SV Gel and PCR Clean-up System, 50 preps Promega Italia Srl Cat# A9281 Deposited data SMP180 dataset Abdul-Gader et al. 96 https://doi.org/10.1093/bioinformatics/ btr234 Pseudomonas Genome Database SciCrunch Registry RRID:SCR_006590, http://www. pseudomonas.com/ Oligonucleotides See Table S3 for Oligonucleotides Recombinant DNA pET28a-TEV-PaCysK2709 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com pET28a-TEV-PaCysK0932 GenScript, Piscataway, NJ, USA RRID:SCR_002891, http://www.genscript. com (Continued on next page) iScience 29, 114304, January 16, 2026 e2



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