Part:BBa_K200014:Design
Opiorphin + EK cleavage site + His Tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
Polyhistidine-tag
Six histidine residues are found upstream of the EK recognition sequence. Since histidine is a charged amino acid, it can be used in affinity purification to obtain high grade opiorphin.
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 sequence.