Difference between revisions of "Part:BBa K1943020:Design"

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===Design Notes===
 
===Design Notes===
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This TetOn expression system contains a SV40 promoter, a puro anti-antibiotics gene, 2A and TetOn 3G promoter. We design primers and do PCR of this TetOn expression system and ligate it to pSB1C3 backbone.
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This year, R-GECO is one of core protein of our project. We find its sequence and synthesis it and ligate it to the plasmid that we transfect to mammalian cells. So we design primers and do PCR of this R-GECO coding sequence and ligate it to pSB1C3 backbone. Besides, there is a R-GECO in biobrick registry, BBa_K881000. However, there are two PstI enzyme cutting sites in BBa_K881000. Thus, we do two site-directed mutagenesis of the two PstI enzyme cutting sites by the degeneracy of condons to make it RCF10 compatible.
  
 
===Source===
 
===Source===

Revision as of 14:00, 16 October 2016


R-GECO


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 914
    Illegal SapI.rc site found at 111
    Illegal SapI.rc site found at 820


Design Notes

This year, R-GECO is one of core protein of our project. We find its sequence and synthesis it and ligate it to the plasmid that we transfect to mammalian cells. So we design primers and do PCR of this R-GECO coding sequence and ligate it to pSB1C3 backbone. Besides, there is a R-GECO in biobrick registry, BBa_K881000. However, there are two PstI enzyme cutting sites in BBa_K881000. Thus, we do two site-directed mutagenesis of the two PstI enzyme cutting sites by the degeneracy of condons to make it RCF10 compatible.

Source

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References