Conjugation

Part:BBa_K1510624:Design

Designed by: Kao Jung Chang   Group: iGEM14_NYMU-Taipei   (2014-10-08)


Truncated lysostaphin coding sequence plus strong promoter and yebf


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 50
    Illegal NheI site found at 73
    Illegal NheI site found at 1092
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 202
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To fit igem part standard, the circuit is constructed in backbone pSB1C3, chloramphenicol with EcoR1 and Xba1 in front of the part, and Spe1, Pst1 behind the part.


Source

The promoter, J23100, comes from the 2014 distributed igem kit. The signal sequence, yebF, encoding signal protein exporting recombinant protein extracellular, is from E.coli K-12. By designing primer, we obtain it together with its naturally occurring ribosomal binding site using PCR and extraction. Finally, the truncated lysostaphin coding sequence is from 2013 igem distribution.

References