Coding sequence for trunctated Lysostaphin fused to His-tagged HlyA
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21INCOMPATIBLE WITH RFCIllegal BglII site found at 1351
- 23COMPATIBLE WITH RFC
- 25INCOMPATIBLE WITH RFCIllegal AgeI site found at 126
- 1000COMPATIBLE WITH RFC
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.
The designed sequence was synthesized de novo by IDT.
2) Waugh, D. (2011). An Overview of Enzymatic Reagents for the Removal of Affinity Tags. Protein Expr Purif., 283-293.