Part:BBa_K5269024

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

It contains SD-sequences(BBa_K5269034), SpyTag, Endo-1,3(4)-b-glucanase(BBa_5269016) and His-Tag 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 SpyCatcher-SpyTag system works when used with Lpp-ompA-SpyCatcher(BBa_K5269026) or INPNC-SpyCatcher(BBa_K5269027). Therefore, Endo-1,3(4)-b-glucanase, which hydrolyzes b-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]