Difference between revisions of "Part:BBa K2812004:Design"

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===Design Notes===
 
===Design Notes===
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Total DNA sequence has been codon optimized for E. coli.  
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The initial starting sequence for truncated lysostaphin was taken from <partinfo>BBa_K748002</partinfo>. The amino acid sequence for HlyA was taken from uniprot (UniProtKB - Q1R2T5 (Q1R2T5_ECOUT)), residue 807-1024.<sup>1</sup> 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, 2011<sup>2</sup> 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''.
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===Source===
 
===Source===

Revision as of 08:45, 16 October 2018


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#sequences