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
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. |