Composite

Part:BBa_K1431201:Design

Designed by: Rifei Chen, Yushan Zhang   Group: iGEM14_SUSTC-Shenzhen   (2014-10-09)
Revision as of 09:51, 17 October 2014 by ChenRifei (Talk | contribs) (→‎Design Notes)

NLS, lead protein into the nucleus


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 66
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 88
    Illegal NgoMIV site found at 107
  • 1000
    COMPATIBLE 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. See the mechanism of BbsI site work below, 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.

References