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. PubMed Article

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