Part:BBa_K358006:Design

lambda lysis cassette with terminator

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
  COMPATIBLE WITH RFC[1000]

Design Notes

We had two steps of PCR to prepare this part.

First, we performed PCR to get the lysis cassette[SRRz] and inserted before double terminator[BBa_B0015].

As the template, we used λ -Hind Ⅲ digest[http://catalog.takara-bio.co.jp/PDFFiles/3403_DS_j.pdf].

Fwd primer:

Rev primer:

Then, we done the point mutation PCR. This "λ -Hind Ⅲ digest" isn't the wt-DNA and there is a mutation on S gene.

By doing this PCR, we prepared wt-SRRz gene.

Fwd primer:

Rev primer:

See details in notebook.

Source

Used λ -Hind digest and BBa_B0015.

References

C. Chang, K. Nam, and R. Young, “S gene expression and the timing of lysis by bacteriophage lambda,” J. Bacteriol., vol. 177, Jun. 1995, pp. 3283-3294.[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC177022/pdf/1773283.pdf]