Coding
TALE2

Part:BBa_K1861601:Design

Designed by: Vladyslav Vyshnevskyi   Group: iGEM15_FAU_Erlangen   (2015-09-18)

TALE that binds the sequence T GTCGTAACAACT


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2176
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1972
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 442
    Illegal AgeI site found at 1149
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This is a component of BBa_K1861060.
As the TALE's repeats proved difficult to synthesize reliably, we used DNAWorks to remove repeats while keeping the same amino acid sequence.
In order to insert the variable TALE domain (VD) into the constant domain (CD) BBa_K1861201, we had to introduce two restriction sites for Esp3I and a spacer between them in the CD, as well as two Esp3I sites in each half of the VD. Thus, we ligated the 5' Esp3I site of the CD with the complementary 5' site of the VD's first half, its 3' site with the VD's second half's 5' site, and VD's second half's 3' site with the CD's 3' site.

Source

The sequence was obtained using the TAL Plasmids Sequence Assembly Tool.

References

http://helixweb.nih.gov/dnaworks/, visited on 2015-09-21
http://web.archive.org/web/20150921225027/http://helixweb.nih.gov/dnaworks/
http://bao.rice.edu/Research/BioinformaticTools/assembleTALSequences.html, visited on 2015-09-20 http://web.archive.org/web/20150920175258/http://bao.rice.edu/Research/BioinformaticTools/assembleTALSequences.html