RNA
Part:BBa_K3044012:Design
Designed by: Catharina Bang Jensen Group: iGEM19_SDU-Denmark (2019-10-07)
sgRNA targeting GFP
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The sgRNA was designed according to the design protocol "CRISPR interference (CRISPRi) for sequence-specific control of gene expression". [1] An important thing to keep ind mind when designing a sgRNA is off-targets. Therefore, the sgRNA sequence was aligned to the core genome of our chassis E coli K12 TOP10 and KG22 and no off-targets were found. Though, off-targets may appear in other bacteria strains.
Source
The sequence of the sgRNA is designed on the basis of the GFP sequence. The target sequence is determined according to the location of PAM sequences in the target sequence, which the Cas protein regcognises.