Difference between revisions of "Part:BBa K2361000"
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In this part the start codon (ATG) starts directly behind the XbaI site and not at the last A of the XbaI site. This may lead to reduced efficiency of expression when combined with RBS's from the iGEM repository, but more importantly this needs to be taken into consideration when creating fusion proteins with this part to ensure that no frame-shifts occur. | In this part the start codon (ATG) starts directly behind the XbaI site and not at the last A of the XbaI site. This may lead to reduced efficiency of expression when combined with RBS's from the iGEM repository, but more importantly this needs to be taken into consideration when creating fusion proteins with this part to ensure that no frame-shifts occur. | ||
| + | ===References=== | ||
[1] Nielsen, A. A., & Voigt, C. A. (2014). Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. Molecular Systems Biology, 10(11), 763. http://doi.org/10.15252/msb.20145735 | [1] Nielsen, A. A., & Voigt, C. A. (2014). Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. Molecular Systems Biology, 10(11), 763. http://doi.org/10.15252/msb.20145735 | ||
Revision as of 08:57, 30 October 2017
spdCas9
This part contains the protein coding sequence for the dCas9 originating from Streptococcus pyogenes. This protein is a catalytically-dead Cas9 variant, which lacks endonuclease activity. It is used for gene repression using CRISPR-interference.
Usage and Biology
Overexpression of dCas9 may cause cytotoxic effects [1], therefore we recommend expressing it using an inducible promotor. In this part the start codon (ATG) starts directly behind the XbaI site and not at the last A of the XbaI site. This may lead to reduced efficiency of expression when combined with RBS's from the iGEM repository, but more importantly this needs to be taken into consideration when creating fusion proteins with this part to ensure that no frame-shifts occur.
References
[1] Nielsen, A. A., & Voigt, C. A. (2014). Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. Molecular Systems Biology, 10(11), 763. http://doi.org/10.15252/msb.20145735
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1100
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3379
Illegal XhoI site found at 4115 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]