Part:BBa_K2812005:Design
Coding sequence for trunctated Lysostaphin with HlyA and His6-tag regulated by T7-promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1389
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 164
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The sequence for the T7 promotor was has been taken from BBa_K525998. 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 807-1024.1 This is the C-terminal signal sequence, the N-terminal catalytic domain responsible for lysis of red blood cells is not included. The thrombin substrate sequence was obtained from Waugh, 20112 and all three components were linked via two flexible GGGGS linkers in between. This amino acid sequence was used to generate a codon optimized sequence for expression in E. coli.
Source
The designed sequence for lysostaphin-thrombin linker-HlyA-His-tag was synthesized de novo by IDT.