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Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity.

Moreb EA, Hoover B, Yaseen A, Valyasevi N, Roecker Z, Menacho-Melgar R, Lynch MD
ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
89951pKDsgRNA-malGContains arabinose-induced lambda Red and a tet-inducible gRNA targeting malG.
89952pKDsgRNA-arsBContains arabinose-induced lambda Red and a tet-inducible gRNA targeting arsB.
89953pKDsgRNA-rpsLContains arabinose-induced lambda Red and a tet-inducible gRNA targeting rpsL.
89954pKDsgRNA-hisGContains arabinose-induced lambda Red and a tet-inducible gRNA targeting hisG.
89955pKDsgRNA-trmIContains arabinose-induced lambda Red and a tet-inducible gRNA targeting trmI.
89956pKDsgRNA-torZContains arabinose-induced lambda Red and a tet-inducible gRNA targeting torZ.
89957pKDsgRNA-rssAContains arabinose-induced lambda Red and a tet-inducible gRNA targeting rssA.
89958pKDsgRNA-yhhTContains arabinose-induced lambda Red and a tet-inducible gRNA targeting yhhT.
89959pKDsgRNA-lpoBContains arabinose-induced lambda Red and a tet-inducible gRNA targeting lpoB.
89960pKDsgRNA-NTContains arabinose-induced lambda Red and a tet-inducible non-targeting gRNA containing BsmBI sites for cloning.
89961pEM-Cas9HF1Modified from pCas9-CR4 (Addgene: 62655) to use the high-fidelity version of Cas9, SpCas9-HF1 (N497A/R661A/Q695A/Q926A) from Kleinstiver et al 2016.
89962pEM-Cas9HF1-recA56Modified from pEM-Cas9HF1 (Addgene ID: 89961) to include constitutive recA56 to block recA-mediated double-strand break repair.
89963pKDsgRNA-rbsKContains arabinose-induced lambda Red and a tet-inducible gRNA targeting rbsK.
89964pKDsgRNA-xylAContains arabinose-induced lambda Red and a tet-inducible gRNA targeting xylA.
89965pKDsgRNA-srlAContains arabinose-induced lambda Red and a tet-inducible gRNA targeting srlA.
102283pSMART-SOS-GFPuvFluorescent sensor of E. coli SOS response
102284pKDsgRNA-pfkBContains arabinose-induced lambda Red and a tet-inducible gRNA targeting pfkB.
102285pEM-Cas9noSsrAModified from pCas9-CR4 (Addgene: 62655) to remove the ssrA tag from Cas9.
102286pKDsgRNA-fsaAContains arabinose-induced lambda Red and a tet-inducible gRNA targeting fsaA.
102288pKDsgRNA-ldhAContains arabinose-induced lambda Red and a tet-inducible gRNA targeting ldhA.
102290pKDsgRNA-adhEContains arabinose-induced lambda Red and a tet-inducible gRNA targeting adhE.
102291pKDsgRNA-ompTContains arabinose-induced lambda Red and a tet-inducible gRNA targeting ompT.
102294pEM-Cas9HF1-indRecA56Modified from pEM-Cas9HF1 (Addgene ID: 89961) to co-express inducible recA56 to block recA-mediated double-strand break repair.

Antibodies from Article