Difference between revisions of "Part:BBa K1189000:Design"

 
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<partinfo>BBa_K1189000 short</partinfo>
 
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===Design Notes===
 
===Design Notes===
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asdsada
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On the iGEM parts registry we found TALA (BBa_K782004) that the Slovenian team synthesized for a previous project. TALEs are very large proteins and take a long time to design and synthesize. Therefore, we decided to use these TALEs to test our system. We also saw this as an opportunity to use and build upon parts made by former iGEM teams. When we sequenced TALA (BBa_K782004), we discovered that it had a small mutated segment. We expected a sequence of AGCAATGGG in the repeat variable di-residue of the second repeat. 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. The TALE also had a kozak sequence at the front of the sequence, we used PCR to remove the kozak sequence at the front of the TALE, so it would not inhibit the expression
  
  

Revision as of 22:36, 27 September 2013

TALE-A with a his tag under a lacI promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2654
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

On the iGEM parts registry we found TALA (BBa_K782004) that the Slovenian team synthesized for a previous project. TALEs are very large proteins and take a long time to design and synthesize. Therefore, we decided to use these TALEs to test our system. We also saw this as an opportunity to use and build upon parts made by former iGEM teams. When we sequenced TALA (BBa_K782004), we discovered that it had a small mutated segment. We expected a sequence of AGCAATGGG in the repeat variable di-residue of the second repeat. 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. The TALE also had a kozak sequence at the front of the sequence, we used PCR to remove the kozak sequence at the front of the TALE, so it would not inhibit the expression


Source

asdasd

References