CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering.

Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH
Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 10.1038/s41587-020-00745-y. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
160729	pSL1521 (pSPIN, pSC101* backbone)	Single-plasmid V. cholerae CAST system. Encodes all proteins, crRNA, donor DNA; non-targeting crRNA has BsaI sites for cloning. Temperature-sensitive pSC101* backbone can be cured by 37 °C incubation.
160730	pSL1142 (pSPIN, pBBR1 backbone)	Single-plasmid V. cholerae CAST system, encodes all proteins, crRNA, and donor DNA. Entry vector encodes non-targeting crRNA, with BsaI sites for spacer cloning. pBBR1 backbone.
160731	pSL1777 (pEffector)	Effector expression for V. cholerae CAST system, encodes all protein and RNA components. Entry vector encodes non-targeting crRNA, with BsaI sites for spacer cloning. pCDF backbone.
160732	pSL1119 (pDonor)	Mini-transposon donor plasmid for V. cholerae CAST system. pUC19 vector backbone.
160733	pSL1749 (pDonor, R*MmeI)	Mini-transposon donor plasmid for V. cholerae CAST system. Mini-transposon has MmeI site in R end for Tn-seq. pUC19 vector backbone.
160734	pSL1765 (pSPIN-R, R*MmeI)	Single-plasmid V. cholerae CAST. Encodes all proteins, crRNA, donor DNA; non-targeting crRNA with BsaI sites. Reverse (R) order minimizes self-targeting; MmeI site for Tn-seq; pBBR1 backbone.
160735	pSL1425 (pSPIN, pCDF backbone)	Single-plasmid V. cholerae CAST system, encodes all proteins, crRNA, and donor DNA. Entry vector encodes non-targeting crRNA and has BsaI sites for spacer cloning. pCDF backbone.
160736	pSL1145 (pSPIN, pBBR1 backbone, R*MmeI)	Single-plasmid V. cholerae CAST, encodes all proteins, crRNA, and donor DNA. Non-targeting crRNA with BsaI sites for spacer cloning. Mini-tn has MmeI site in R end for Tn-seq. pBBR1 backbone.