Part:BBa_K4370012:Design
sgRNA_gpm_STREPTO
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This sequence has been designed to target different strains such as S. ambofaciens ATCC 23877, S. bottropensis ATCC 25435, and S. venezuelae ATCC 10712. The part is also designed to be cloned between NcoI (CCATGG) and SnaBI (TACGTA) sites into the pCRISPR-dCas9 plasmid published by Tong and collaborators (ACS Synthetic Biology, 2015). This plasmid encodes a dead version of Cas9 that can be used to silence gene expression at specific loci. Twenty nucl. (TCTGGGCCTTGTCCTTGCCC) are perfect the gpm gene, the following nucleotides (GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCT) correspond to the gRNA scaffold.
Please have a look to our CRISPRi tuto to learn more about the way to design sgRNA for gene expression silencing in bacteria.
Source
The DNA has been synthetized.