Part:BBa_K1189005:Design

TALEB Target ([B])

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 213
  Illegal AgeI site found at 325
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

We designed primers that would produce these TALE B target sequence in a KAPA PCR reaction. Our target sequences were then ligated into RFP generator, in a pSB1C3 backbone. We expected a sequence of AGCAATGGG in the repeat variable di-residue of the second repeat of TALB, however, the sequence was actually TCCCACGAC. This meant that the required target sequence at this position was a C, and not a T, as the parts registry web page indicates. Therefore, this part has the updated TALB target sequence with C instead of the T.

Source

The target sequence was amplified with primers from the genome of E. coli using polymerase chain reaction.

References

Bogdanove, A. J., Schornack, S., & Lahaye, T. (2010). TAL effectors: finding plant genes for disease and defense. Current Opinion in Plant Biology, 13(4), 394–401. doi:10.1016/j.pbi.2010.04.010

Mussolino, C., & Cathomen, T. (2012). TALE nucleases: tailored genome engineering made easy. Current Opinion in Biotechnology, 23(5), 644–50. doi:10.1016/j.copbio.2012.01.013