Part:BBa_K3941005

pET29b+EGII

BBa_K3941005 is a composite part that we used in our expression plasmids (pET29b). We obtain EG2 wild type with this sequence.

This composite part contains 4 parts:

- T7 Promoter (BBa_K3941000)
- LacO (BBa_K1624002)
- EGII (BBa_K3941001)
- T7 Terminator

Figure 1: T7 promoter, Lac Operon, Codon optimized EGII, 6XHis and terminator

Codon OptimizedEGII

This composite part originated from a codon-optimized (for E. coli DH5⍺) version of the endoglucanase (EG) gene that cleaves the internal beta-1,4-glycosidic bonds in cellulose. We optimized the sequence for expression and added a 6XHis at the end. This part contains a mutation to form a different amino acid than wild type. We changed the Cystine from the 99th amino acid to Valine.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 22
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 93
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 22
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 22
  
• 1000
  COMPATIBLE WITH RFC[1000]