Coding

Part:BBa_K200013:Design

Designed by: James Field   Group: iGEM09_Imperial College London   (2009-10-13)
Revision as of 15:35, 18 October 2009 by JamesField (Talk | contribs) (→‎Design Notes)

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

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.

File:Https://static.igem.org/mediawiki/2009/a/a2/Opiorphin.png

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