Composite

Part:BBa_K5269022

Designed by: Yuichiro Nakai   Group: iGEM24_TokyoTech   (2024-09-28)
Revision as of 14:25, 28 September 2024 by Registry (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


SpyTag-GH19Chitinase-His-Tag and BsmB I cut site(for GGA)

This sequence contains SD sequence, SpyTag (BBa_K5269017), GH19 (BBa_K5269015) and 6xHis sequence. In addition, 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 SpyTag/SpyCatcher system works when used with Lpp-ompA-SpyCatcher(BBa_K5269026) or INPNC-SpyCatcher(BBa_K5269027). Therefore, GH19, 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 20
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 356
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 885
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 20
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 20
    Illegal NgoMIV site found at 686
    Illegal AgeI site found at 641
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None