Composite

Part:BBa_K5269024

Designed by: Yuichiro Nakai   Group: iGEM24_TokyoTech   (2024-09-28)
Revision as of 17:34, 28 September 2024 by Nakaiyuichiro (Talk | contribs)


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

It contains SD sequences(BBa_K5269034), SpyTag, Endo-1,3(4)-beta-glucanase(BBa_5269016) and 6xHis sequences. 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, Endo-1,3(4)-beta-glucanase, which hydrolyzes beta-D-glucan 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
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 235
    Illegal NotI site found at 1603
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2845
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 627
    Illegal NgoMIV site found at 1252
    Illegal NgoMIV site found at 1995
    Illegal NgoMIV site found at 2620
    Illegal AgeI site found at 385
    Illegal AgeI site found at 1753
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None