Part:BBa_K3941004

pET29b+CelAB

BBa_K3941004 is a sequence that we used in our expression plasmids (pET29b). We produce CelAB via this specific sequence.

We ordered these plasmids from IDT.

This composite part is made out of 4 different parts:

- T7 Promoter (BBa_K3633015)
- LacO (BBa_K1624002)
- CelAB (BBa_K3941000)
- T7 Terminator

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

Codon Optimized CelAB

This composite part uses a codon-optimized (for E. coli DH5⍺) version of the CelAB's catalytic region's sequence. We optimized the sequence for expression and added a 6XHis at the end. It is created to produce an endo-1, 4-β-D glucanase protein to degrade cellulose and its derivatives.

We took the catalytic region of the original CelAB sequence to use in our experiments and later on optimized it to use in E. coli DH5⍺ bacteria. We also used E. coli BL21 to express our plasmids.

To harvest our proteins, we added a His-tag (6XHis) at the end of our AA sequence.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 51
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 930
  Illegal XhoI site found at 972
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 51
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 51
  
• 1000
  COMPATIBLE WITH RFC[1000]