Difference between revisions of "Part:BBa K3941006"
 
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<partinfo>BBa_K3941006 short</partinfo>
 
<partinfo>BBa_K3941006 short</partinfo>
  
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BBa_K3941006 is a composite part that we used in our expression plasmids (pET29b). We obtain EG2 wild type with this sequence.
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This composite part contains 4 parts:
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- T7 Promoter (BBa_K3941000)<br>
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- LacO (BBa_K1624002)<br>
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- EGII (C99V) (BBa_K3941002)<br>
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- T7 Terminator
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https://2021.igem.org/wiki/images/thumb/3/3e/T--Saint_Joseph--EGII-C99V-Whole.png/800px-T--Saint_Joseph--EGII-C99V-Whole.png
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<font size="-2"><b>Figure 1:</b> T7 promoter, Lac Operon, Codon optimized EGII, 6XHis and terminator</font>
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<font size="+1"><b>Codon Optimized EGII (C99V)</b></font>
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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.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3941006 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3941005 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3941006 parameters</partinfo>
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<partinfo>BBa_K3941005 parameters</partinfo>
 
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Latest revision as of 11:42, 19 October 2021


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


800px-T--Saint_Joseph--EGII-C99V-Whole.png

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]