Composite
Part:BBa_K1431201:Design
Designed by: Rifei Chen, Yushan Zhang Group: iGEM14_SUSTC-Shenzhen (2014-10-09)
NLS, lead protein into the nucleus
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 66
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 88
Illegal NgoMIV site found at 107 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We have anther kind of NLS but only with one side. We chose two NLS for it may increase the probability of protein import into the cell nucleus. As for BbsI site, we have a series same design sequence. You can get how BbsI site work by google, and the reason why we designed that because we want to make a seamless gather between NLS and protein or any triple nucleotides. That depend on which will make the highest efficiency. Sorry for the limited time, we do not test which will be the highest efficiency.
Source
These two NLS sequence was from two sides in Cas9 sequence, plasmid px330, zhangfeng lab. For the too length and repeat A&G bases, we completely synthesis the sequence and add two BbsI site specially.