Part:BBa_K1189033:Design

EHEC TALE2 (binds to a 19bp region of stx2 gene)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2404
  Illegal BamHI site found at 2539
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

A KasI restriction cut site has been incorporated in the end of the TALE, so that part can easily be used to replace the TALEs in our other constructs.

Source

This part was commercially synthesized using four IDT G-Blocks and a linear pSB1C3 PCR product. SOE PCR (Heckman & Pease, 2007) was used to assemble the four gene blocks in two steps whilst Gibson assembly was used to place the fragment (Figure 2) into pSB1C3.

References

Heckman, K. L., & Pease, L. R. (2007). Gene splicing and mutagenesis by PCR-driven overlap extension. Nature protocols, 2(4), 924-932.

Meckler, J.F., Bhakta, M.S., Kim, M.S., Ovadia, R., Habrian, C.H., Zykovich, A., ... Baldwin, E.P. (2013). Quantitative analysis of TALE-DNA interactions suggests polarity effects. Nucleic Acids Research, 41(7), 4118–28. doi:10.1093/nar/gkt085

Perna, N. T., Plunkett, G., Burland, V., Mau, B., Glasner, J. D., Rose, D. J., ... Blattner, F. R. (2000) Genome sequence of enterohaemorrhagic Escherichia coli O157:H7. Letters to Nature. 409, 529-533. doi:10.1038/35054089