Coding

Part:BBa_K523010

Designed by: Mun Ching Lee, Sylvia Ispasanie, Allan Crossman   Group: iGEM11_Edinburgh   (2011-08-04)

β-glucosidase for BioSandwich

This is a modified version of BBa_K392008 for use with the BioSandwich protocol of Edinburgh 2011. However, since the published sequence for K392008 has significant deviations from reality, the expected sequence for this new part does not match it precisely. Also, we strongly believe that it is the 2nd ATG of BBa_K392008 which is the true start of the coding sequence; therefore everything upstream of that was omitted in this new part.

When used to make fusion proteins, BioSandwich assembly requires an in-frame BglII site (formed by the last 2 bases of the RFC10 prefix and the first 4 bases at the start of this BioBrick) and an in-frame SpeI site (present in the RFC10 suffix). Because the SpeI site is out of frame, two extra "G" bases have been added at the end. When combined with the T at the start of the RFC10 suffix, they code for a glycine residue and correct the frameshift.

During the creation of this part, a point mutation arose converting an arginine into a cysteine.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 863
    Illegal XhoI site found at 1097
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 350
    Illegal AgeI site found at 246
    Illegal AgeI site found at 441
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 223


[edit]
Categories
Parameters
None