Difference between revisions of "Part:BBa K3941005"

Revision as of 21:05, 17 October 2021

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]

@@ Line 7: / Line 7: @@
 This composite part contains 4 parts:
-- T7 Promoter (BBa_K3941000)
+- T7 Promoter (BBa_K3941000)<br>
+- LacO (BBa_K1624002)<br>
-- LacO (BBa_K1624002)
+- EGII (BBa_K3941001)<br>
-- EGII (BBa_K3941001)
 - T7 Terminator
 https://2021.igem.org/wiki/images/thumb/c/cd/T--Saint_Joseph--EGII-Whole.png/800px-T--Saint_Joseph--EGII-Whole.png
+<font size="-2"><b>Figure 1:</b> T7 promoter, Lac Operon, Codon optimized EGII, 6XHis and terminator</font>
-<font size="+1"><b>EGII</b></font>
+<font size="+1"><b>Codon OptimizedEGII</b></font>
 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.