Difference between revisions of "Part:BBa K200013:Design"

 
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<partinfo>BBa_K200013 short</partinfo>
 
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===Design Notes===
 
===Design Notes===
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Since this is a pentapeptide that does not start with methionine, we had to design on a cleavable linker.
 
  
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==Cleavable Linker==
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[[Image:Opiorphin.png|left|400px]]Since polypeptides can only be biologically synthesised if they start with the amino acid methionine, it has been necessary to design a cleavable peptide sequence starting with methionine. This sequence is found upstream of opiorphin and is enzymatically cleaved following incubation with the enzyme enterokinase. Enterokinase is a highly specific serine protease that cleaves after the recognition sequence Aspartic Acid - Aspartic Acid - Aspartic Acid -Aspartic Acid – Lysine.
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The methionine is shown in RED, the protease recognition site is shown in BLUE and opiorphin is shown in GREEN. The RED & BLUE sections make up the removable linker region which is cleaved by the serine protease enterokinase.
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==Stop Codon==
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A double stop codon has been incorporated into the sequence to ensure efficient termination.
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==Codon Choice==
  
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The sequence has been optimised for expression in <i>E.coli</i>.
  
 
===Source===
 
===Source===

Latest revision as of 15:37, 18 October 2009

Opiorphin + EK cleavage site


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Cleavable Linker

Opiorphin.png
Since polypeptides can only be biologically synthesised if they start with the amino acid methionine, it has been necessary to design a cleavable peptide sequence starting with methionine. This sequence is found upstream of opiorphin and is enzymatically cleaved following incubation with the enzyme enterokinase. Enterokinase is a highly specific serine protease that cleaves after the recognition sequence Aspartic Acid - Aspartic Acid - Aspartic Acid -Aspartic Acid – Lysine.

The methionine is shown in RED, the protease recognition site is shown in BLUE and opiorphin is shown in GREEN. The RED & BLUE sections make up the removable linker region which is cleaved by the serine protease enterokinase.

Stop Codon

A double stop codon has been incorporated into the sequence to ensure efficient termination.


Codon Choice

The sequence has been optimised for expression in E.coli.

Source

Synthetic part.

References