Simultaneous multi-site editing of individual genomes using retron arrays.
Gonzalez-Delgado A, Lopez SC, Rojas-Montero M, Fishman CB, Shipman SL
bioRxiv [Preprint]. 2023 Jul 17:2023.07.17.549397. doi: 10.1101/2023.07.17.549397.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 219497 | pAGD236 | pCDF-Duet deriviative with sfGFP flanked by 2 BsaI sites downstream of the first T7 promoter and independent msr and RT downstream of the second T7 promoter |
| 219498 | pAGD248 | Temperature sensitive replication (repA101ts) backbone encoding the retron recombineering operon (ncRNA, RT, CspRecT, mutLE32K) |
| 219499 | pAGD335 | pAGD248 derivative with the retron Eco1 msr, sfGFP flanked by 2 BsaI sites , and the rest of the retron recombineering operon (Eco1 RT, CspRect, MutL-E32K) downstream of the pBad promoter |
| 219500 | pSCL390 | Integrating inducible cassette (Cas9-P2A-Csy4 and Eco1 RT) for yeast overexpression |
| 219501 | pSCL391 | Express Ade2 and Faa1 editrons from a single Gal7 promoter, separated by a Csy4 recognition sequence |
| 219502 | pSCL452 | Golden Gate entry vector for library cloning of msd-sgRNA arrays: GAL7p-Csy4-PaqCI-PaqCI-GAL7t // SNR52p-msr-SUP4t |
| 219503 | pSCL475 | Express Ade2, Can1 and Faa1 retron msd donor-gRNAs from a single Gal7 promoter, separated by a Csy4 recognition sequence, with an msr expressed in trans |
| 219504 | pSCL758 | Express Eco3RT from a CAG promoter; also express an msr and HEK3, FANCF and EMX1 retron msd donor-gRNAs from a single H1 promoter, separated by tRNA-Cys-GCA sequences |