Part:BBa_K3941006

pET29b+EGII (C99V)

BBa_K3941006 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 (C99V) (BBa_K3941002)
- T7 Terminator

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

Codon Optimized EGII (C99V)

This part is created based on only the coding sequence of Trichoderma reesei endoglucanase II gene by excluding introns and non-coding exon sites. Signal sequence is also removed. Codon optimization has carried out to increase production in Escherichia coli bacterium. His-tag was added to 3' end of the sequence to help purification of protein product. A single aminoacid mutation has been studied as Cysteine acid aminoacid in 99 aminoacid position is changed with Valine aminoacid.

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]