Composite

Part:BBa_K3941007

Designed by: Can Bora Ciner   Group: iGEM21_Saint_Joseph   (2021-10-19)
Revision as of 11:40, 19 October 2021 by Boraciner (Talk | contribs)


pET29b+EGII (D185S)

BBa_K3941007 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


800px-T--Saint_Joseph--EGII-Whole.png

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


Codon Optimized EGII (D185)

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 Aspartic acid aminoacid in 185 aminoacid position is changed with Serine 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]


[edit]
Categories
Parameters
None