Coding

Part:BBa_K4645008

Designed by: Meng Sun   Group: iGEM23_HZAU-China   (2023-10-10)
Revision as of 08:50, 12 October 2023 by Felicitysm (Talk | contribs)

A chimeric lysozyme with secretory ability.

ClyR is a chimeric lysin, comprising a cysteine, histidine-dependent amidohydrolase/peptidase catalytic domain from PlyC lysin, and the cell-wall binding domain from PlySs2 lysin. The signal peptide OmpA enables ClyR to be secreted into the periplasmic space of bacteria.


Result

We used homologous recombination in which the OmpA signal peptide was added to the N-terminus of ClyR and transformed into E.coli DH5α competent cells and incubated overnight at 37℃. Below is a plot of colony PCR results.


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Figure 1. Colony PCR assay of the OmpA-ClyR fragment.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 816
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 184
    Illegal AgeI site found at 268
    Illegal AgeI site found at 583
    Illegal AgeI site found at 766
  • 1000
    COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None