Difference between revisions of "Part:BBa K2915285:Design"

 
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===References===
 
===References===
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Cermak T, et al. Efficient design and assembly of custom TALEs and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res 2011 Sep 1;39(17):7879.

Latest revision as of 20:31, 19 October 2019


IPTG inducible promoter (T7) with RBS TAL3 6-His-Tag with TEV site


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 639
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 698
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 272
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 252


Design Notes

We add a 6Hig-Tag and a TeV cleavage site into the "TAL3" sequence which allows us to purify our proteins. We optimized the codons to be able to produce these proteins into the K12 E.coli strain. To perform the production f TAL3 in a K12 E.coli strain, we added a T7 promotor and a RBS into the pSB1C3 plasmide,which is the regulator.. This biobrick is in RFC 10 standards with: The prefixe (with ATG in frame): 5' GAATTC GCGGCCGC T TCTAGA TG GCCGGC and the suffixe (contain Stop in frame): ACCGGT TAAT ACTAGT A GCGGCCG CTGCAG 3'


Source

Nucleics acids

References

Cermak T, et al. Efficient design and assembly of custom TALEs and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res 2011 Sep 1;39(17):7879.