Composite

Part:BBa_K3585007:Design

Designed by: Kong Yangyang   Group: iGEM20_Shanghai_HS_United   (2020-10-14)


pJ23100-butyrate-J23200-lacY


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1240
    Illegal NheI site found at 8195
    Illegal NheI site found at 8218
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 6272
    Illegal BglII site found at 7408
    Illegal BglII site found at 7549
    Illegal XhoI site found at 1245
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 9008
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In order to meet the TypeIIS standard, we introduced mutations of Asp50Asp and Leu137Leu in the ptb and crt domains respectively to remove the BsaI restriction enzyme site.


Source

The promoter sequence comes from Part:BBa_J23100. The biosynthetic gene cluster comes from Part:BBa_K3585001. The sequence of lacY comes from the genomic sequence of E. coli Nissle 1917.

References