Coding
Part:BBa_K2812004:Design
Designed by: Guido Oerlemans, Maxime van den Oetelaar and Mariska Brüls Group: iGEM18_TU-Eindhoven (2018-10-03)
Coding sequence for trunctated Lysostaphin fused to His-tagged HlyA
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1351
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 126
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Total DNA sequence has been codon optimized for E. coli.
Source
The initial starting sequence for truncated lysostaphin was taken from BBa_K748002. The amino acid sequence for HlyA was taken from uniprot (UniProtKB - Q1R2T5 (Q1R2T5_ECOUT)), residue 807-1024. This is the C-terminal signal sequence, the N-terminal catalytic domain responsible for lysis of red blood cells is not included. The thrombin linker was obtained from uniprot. This amino acid sequence was used to generate a codon optimized sequence for expression in E. coli.