Difference between revisions of "Part:BBa K3941004"
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BBa_K3941004 is a sequence that we used in our expression plasmids (pET29b). We produce CelAB via this specific sequence. | BBa_K3941004 is a sequence that we used in our expression plasmids (pET29b). We produce CelAB via this specific sequence. | ||
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| + | We ordered these plasmids from IDT. | ||
This composite part is made out of 4 different parts: | This composite part is made out of 4 different parts: | ||
| − | - T7 Promoter ( | + | |
| − | - LacO (BBa_K1624002) | + | - T7 Promoter (BBa_K3633015)<br> |
| − | - CelAB (BBa_K3941000) | + | - LacO (BBa_K1624002)<br> |
| + | - CelAB (BBa_K3941000)<br> | ||
- T7 Terminator | - T7 Terminator | ||
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| − | <font size="-2"><b>Figure 1:</b> T7 promoter | + | <font size="-2"><b>Figure 1:</b> T7 promoter, Codon optimized CelAB, 6XHis, terminator and Lac Operon</font> |
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| + | <font size="+2"><b>Codon Optimized CelAB</b></font> | ||
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| + | This composite part uses a codon-optimized (for <i>E. coli</i> 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 <i>E. coli</i> DH5⍺ bacteria. We also used <i>E. coli</i> BL21 to express our plasmids. | ||
| + | |||
| + | To harvest our proteins, we added a His-tag (6XHis) at the end of our AA sequence. | ||
Latest revision as of 21:08, 17 October 2021
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
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 51
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 930
Illegal XhoI site found at 972 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 51
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 51
- 1000COMPATIBLE WITH RFC[1000]