Composite

Part:BBa_K4630117:Design

Designed by: Li Yutong   Group: iGEM23_WHU-China   (2023-10-11)


Xbal-stgframe-Spel-BamHI-selfRecombination


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Firstly, XbaI-stgRNA-1-SpeI-BamHI-selfRecombination is designed by Information Technology and it has relatively high accuracy in achieving the function of self-targeting guidance. We design N20 fragments through the N20 knockout database, and we improve the N20 database through the wet experiment data to form a mutually reinforcing process Secondly, we use enzyme-cutting and enzyme-linking techniques. Among the restriction enzyme cutting sites, XbaI and SpeI executive cutting function to knock out the targeting fragment; the stgRNA-2 fragment will be inserted between sites SpeI and BamHI.Thirdly, we use the restoration system--Lambda system, which has high fidelity repair function.


Source

complete synthesis

References