Composite

Part:BBa_K5269021

Designed by: Yuichiro Nakai   Group: iGEM24_TokyoTech   (2024-09-28)
Revision as of 08:21, 30 September 2024 by Nakaiyuichiro (Talk | contribs)

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SpyTag-ChitinaseC-His-Tag and BsmB I cut site(for GGA)

This sequence contains SD-sequence(BBa_K5269034), SpyTag, ChitinaseC (BBa_K5269014) and His-Tag. The BsmB I cleavage sites at both ends of the sequence allow for Golden Gate Assembly to the Golden Gate Assembly Site (BBa_K5269020) and T7 promoter-shRNA-T7 terminator-Golden Gate Assembly Site(BBa_K5269025). The SpyCatcher-SpyTag system works when used with Lpp-OmpA-SpyCatcher(BBa_K5269026) or INPNC-SpyCatcher(BBa_K5269027). Therefore, ChitinaseC, which hydrolyzes chitin contained in the target bacterial membrane, is expected to be stably present on the Membrane Vesicle, which will facilitate the introduction of shRNA into the target fungus cells.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 7
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1022
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 7
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 7
    Illegal NgoMIV site found at 150
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None