Generator

Part:BBa_K358010:Design

Designed by: Ken Kajita   Group: iGEM10_Kyoto   (2010-09-19)
Revision as of 11:44, 23 October 2010 by Tako (Talk | contribs) (→‎Design Notes)

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anti-killer gene with GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 962


Design Notes

K358010.jpg

We performed three steps of PCR to prepare this part.

First, we got S gene from lambda phage DNA: λ -Hind Ⅲ digest, and inserted into the plasmid, BBa_E0840.

Then, performed the point mutation PCR, the same to the point mutation PCR on SRRz.

Finally, we deleted a transmembrane domain, TMD1, and constructed the anti-killergene: SΔTMD1.

Source

λ -Hind Ⅲ digest (product of TaKaRa Bio) and BBa_E0240

References

R. White, T. A. T. Tran, C. A. Dankenbring, J. Deaton, and R. Young, “The N-terminal transmembrane domain of lambda S is required for holin but not antiholin function,” Journal of Bacteriology, vol. 192, no. 3, pp. 725-733, Feb. 2010.