Composite
Part:BBa_K1323002
Designed by: Julia Manalil Group: iGEM14_Waterloo (2014-10-07)
dCas9: Expression cassette under a xylose inducible promoter
dCas9 or ‘dead’ cas9 is a mutated version of Cas9 that has lost its endonuclease activity - it no longer cuts double stranded DNA, instead simply binding to it. This means it can function as a repressor after complexing with sgRNA and the sgRNA-target site. dCas9 contains mutations in the RuvC1 and HNH nuclease domains; these changes are what bestow this modified Cas9 with it’s ability to bind DNA instead of cleaving it (Lei S. Qi, 2013). It has a Xylose Inducible Promoter (BBa_K1323014) and a sodA RBS (BBa_K1323022). The illegal sites have been removed, and the sequence was codon optimized for expression in S. epidermidis using JCat software.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1476
Illegal BamHI site found at 4894 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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Categories
Parameters
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