Difference between revisions of "Part:BBa K1189005:Design"

 
 
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__NOTOC__
 
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<partinfo>BBa_K1189005 short</partinfo>
 
<partinfo>BBa_K1189005 short</partinfo>
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===Design Notes===
 
===Design Notes===
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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.
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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.
  
  
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===Source===
 
===Source===
  
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The target sequence was amplified with primers from the genome of <i>E. coli</i> using polymerase chain reaction.
  
 
===References===
 
===References===
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<li>Bogdanove, A. J., Schornack, S., & Lahaye, T. (2010). TAL effectors: finding plant genes for disease and defense. <i>Current Opinion in Plant Biology, 13</i>(4), 394–401. doi:10.1016/j.pbi.2010.04.010
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<li>Mussolino, C., & Cathomen, T. (2012). TALE nucleases: tailored genome engineering made easy. <i>Current Opinion in Biotechnology, 23</i>(5), 644–50. doi:10.1016/j.copbio.2012.01.013

Latest revision as of 01:39, 26 October 2013

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