Part:BBa_K523011

Endoglucanase for BioSandwich

This is a modified version of BBa_K118023 for use with the BioSandwich protocol of Edinburgh 2011.

When used to make fusion proteins, 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 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.

The signal peptide of the endoglucanase (amino acids 1 - 31) has been removed, since this part is intended for fusion to INP or other display proteins. Nevertheless a start codon is included so this part can also be used as a control. There is no stop codon.

Some codons at the start were changed to avoid primer secondary structure.

Usage and Biology

This is an endoglucanase from Cellulomonas fimi. It degrades cellulose chains by cutting them in the middle, producing smaller chains.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 1092
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 141
  Illegal XhoI site found at 503
  Illegal XhoI site found at 752
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 283
  Illegal NgoMIV site found at 1208
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 187