Coding

Part:BBa_K523009

Designed by: Sylvia Ispasanie, Mun Ching Lee   Group: iGEM11_Edinburgh   (2011-08-04)
Revision as of 11:58, 10 August 2011 by Allancrossman (Talk | contribs)

Exoglucanase for BioSandwich

This is a modified version of BBa_K118022 for use with the BioSandwich protocol of Edinburgh 2011.

BioSandwich assembly uses 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 bases ("TC") have been added at the end. When combined with the T at the start of the RFC10 suffix, they code for a serine residue and correct the frameshift.

The signal peptide of the exoglucanase (amino acids 1 - 42) has been removed, since this part is intended for fusion to INP or other display proteins.

A non-synonymous change (see figure) was introduced by PCR.

Usage and Biology

This is an exoglucanase from Cellulomonas fimi. It degrades cellulose by chewing at the end of a cellulose chain to produce cellobiose sugar.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 405
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 38
    Illegal NgoMIV site found at 411
    Illegal NgoMIV site found at 913
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 458
    Illegal SapI.rc site found at 541


[edit]
Categories
Parameters
None