Part:BBa_K2812004:Design

Coding sequence for trunctated Lysostaphin fused to His-tagged HlyA

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1351
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 126
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

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 was synthesized de novo by IDT.

References

1) https://www.uniprot.org/uniprot/Q1R2T5

2) Waugh, D. (2011). An Overview of Enzymatic Reagents for the Removal of Affinity Tags. Protein Expr Purif., 283-293.