A set of genetic tools for use in Clostridioides difficile and related species.

Fisher H, de Lussy Kubisa L, Jakhmola A, Walker E, Kirk JA, Oatley P, Chaudhuri RR, Douce GR, Ormsby MJ, Fagan RP
Microbiology (Reading). 2026 Feb;172(2). doi: 10.1099/mic.0.001665. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
245126	pLLK007	Control plasmid based on pAF259 (Addgene #105494). Contains a promoterless luciferase-encoding bitluc.
245127	pEW001	Based on pAF259 but with bitluc expression driven by the C. saccharoperbutylacetonicum bdh promoter.
245128	pEW002	Based on pAF259 but with bitluc expression driven by the C. saccharoperbutylacetonicum Cspa_135p00010 promoter.
245129	pEW003	Based on pAF259 but with bitluc expression driven by the C. saccharoperbutylacetonicum flgB promoter.
245130	pEW004	Based on pAF259 but with bitluc expression driven by the C. saccharoperbutylacetonicum Cspa_RS22190 promoter.
245131	pEW005	Based on pAF259 but with bitluc expression driven by the C. saccharoperbutylacetonicum rpoD promoter.
245132	pEW007	Based on pAF259 but with bitluc expression driven by the C. saccharoperbutylacetonicum Cspa_135p00700 promoter.
245133	pLLK010	Based on pAF259 but with bitluc expression driven by the C. difficile minimal slpA promoter.
245134	pLLK011	Based on pAF259 but with bitluc expression driven by the C. difficile minimal slpA promoter with an additional 37bp upstream.
245135	pLLK012	Based on pAF259 but with bitluc expression driven by the C. difficile minimal slpA promoter with an additional 332bp upstream.
245136	pJAK012	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding an N-term StrepTag II
245137	pJAK013	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding an N-term SNAP tag
245138	pJAK014	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding a C-term SNAP tag
245139	pJAK032	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding an N-term CLIP tag
245140	pJAK033	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding a C-term CLIP tag
245141	pHF008	Expression plasmid carrying the C. sporogenes csxA gene driven by its native promoter and with a C-term mCherry fusion
245142	pPOE025	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding a C-term 3xHA tag
245143	pPOE031	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding a C-term 3xHis tag
245144	pJAK081	Knock-in plasmid for insertion of DNA sequences between CD0188 (pyrE) and CD0189 in the C. difficile R20291 genome.
245145	pJAK122	Expression plasmid with the constitutive Pcwp2 promoter driving expression of mCherry.
245146	pJAK124	pJAK081 carrying Pcwp2-mCherry
245147	pJAK125	pJAK081 carrying Pcwp2-mScarlet-I
245148	pJAK139	Expression plasmid with the constitutive Pcwp2 promoter driving expression of the C. difficile secA2 gene and adding a C-term mCherry.
245149	pJAK201	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 1 (AAGTCCTCG)
245150	pJAK202	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 2 (TCTTGACCG)
245151	pJAK203	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 3 (AACAACACC)
245152	pJAK204	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 4 (AACAGGTGG)
245153	pJAK205	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 5 (ACCGATTAG)
245154	pJAK206	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 6 (CACTTATGC)
245155	pJAK207	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 7 (CCTCCAACT)
245156	pJAK208	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 8 (CGAGGACAT)
245157	pJAK209	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 9 (CTGGTTCTA)
245158	pJAK210	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 10 (GGATGTTGG)
245159	pJAK211	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 11 (GTCACCAGT)
245160	pJAK212	pJAK081-based knock-in plasmid with a 218 bp insertion containing 9 bp sequencing Barcode 12 (AGTGTAACG)
248191	pEW008	Based on pAF259 but with bitluc expression driven by the C. saccharoperbutylacetonicum hsp promoter.