Difference between revisions of "Part:BBa K1638018:Design"

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<partinfo>BBa_K1638018 short</partinfo>
 
<partinfo>BBa_K1638018 short</partinfo>

Revision as of 09:57, 18 September 2015

hTrx-based scaffold fused to T18 through a flexible linker


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 624
    Illegal XhoI site found at 753
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 165
    Illegal NgoMIV site found at 575
    Illegal AgeI site found at 381
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was synthesized by IDT as a gBlock gene fragment.


Library

  • Choose the size of your library. Typically the length of the variable peptide loop is between 10 and 20 amino acids.
  • Dependent on the size your variable peptide loop, you can for example contruct the library as following: ATATGCGCTCGAG(NNK)xGAGCTCATTACCG (where N represent A, T, G or C and K represent G or C). The NNK codon minimizes the number of stop codons. The XhoI restriction included allows insertion of library into the scaffold.
  • Order the library as duplex-DNA
  • When delivered, cut the library and scaffold with XhoI restriction enzyme and ligate.

Important note: This part cannot be used in the standard backbones pSB1C3 or pSB1K3, as they include a xhoI restriction site. Instead we recommend using the pSB1A2 backbone

Source

pUT18C Human thioredoxin sequence: UniProt (AC: P10599) Intein sequence: pTYB21 (IMPACT kit)

References