Synonymous edits in the Escherichia coli genome have substantial and condition-dependent effects on fitness.
Yang DD, Rusch LM, Widney KA, Morgenthaler AB, Copley SD
Proc Natl Acad Sci U S A. 2024 Jan 30;121(5):e2316834121. doi: 10.1073/pnas.2316834121. Epub 2024 Jan 22.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 182958 | pDY118A | pSC101 ori, chl34 resistant. Cas9 induced at 0.4 μg/ml anhydrotetracycline, recombineering proteins induced by a 15 min heat-shock at 42°C, I-Scel (induced by 0.1 mM IPTG) is to cleave the pDonor2. |
| 182959 | pDY279 | Str resistant,CoE1* ori. CRISPR/Cas9 gRNA under constitutive J23119 promoter. gRNA targeting E. coli ptsI codon 2~9. HRT has synonymous mutations in ptsI. (for 1st round editing) |
| 182960 | pDY281 | Amp resistant, CoE1* ori. CRISPR/Cas9 induced by AHTc, gRNA under constitutive J23119 promoter. gRNA targeting E. coli ptsI codon 2~9. HRT has synonymous mutations of ptsI. (for 2nd round editing) |
| 182962 | pDY380 | Amp-resistant, low copy (p15A ori), E. coli 5'ptsI transcriptionally fused with GFP. |
| 182963 | pDY390 | Amp-resistant, low copy (p15A ori), E. coli ptsI transcriptionally fused with GFP. |