Difference between revisions of "Part:BBa K1189001:Design"
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TALB (BBa_K782006) was from parts registry.
 
TALB (BBa_K782006) was from parts registry.
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The new TALB gene without the Kozak sequence was amplified from the genome of <i>E. coli<i/> using polymerase chain reaction.
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The new TALB gene without the Kozak sequence was amplified from the genome of <i>E. coli</i> using polymerase chain reaction.
  
 
===References===
 
===References===

Revision as of 22:45, 27 September 2013

TALE-B with a his 6 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 TALB (BBa_K782006) 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 TALB (BBa_K782006), 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

TALB (BBa_K782006) was from parts registry. The new TALB gene without the Kozak sequence was amplified from the genome of E. coli using polymerase chain reaction.

References