Part:BBa_K1189033:Design
EHEC TALE2 (binds to a 19bp region of stx2 gene)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2404
Illegal BamHI site found at 2539 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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
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
Heckman, K. L., & Pease, L. R. (2007). Gene splicing and mutagenesis by PCR-driven overlap extension. Nature protocols, 2(4), 924-932.