Coding

Part:BBa_K1603003:Design

Designed by: John Hellgren   Group: iGEM15_Chalmers-Gothenburg   (2015-09-18)
Revision as of 13:42, 18 September 2015 by Jannie (Talk | contribs) (→‎Design Notes)


RecA - optimized for yeast


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 260
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1116
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1032


Design Notes

Synthetic gene ordered from IDT.

Codon optimized for Saccharomyces cerevisiae and fused with a nuclear localization signal (NLS) for in vivo DNA repair. Also fused with a His-tag, allowing nickel based purification prior in vitro DNA repair


Primers for amplification with BioBrick Prefix in the FW primer and Suffix in the RV primer:

FW:GCTTCTAGATGTCTCATAGACATCACCATCAC

RV:AGCCTGCAGCGGCCGCTACTAGTATTAAGCTTCTGCGGCTTCA


The primers were designed to remove the EcoRI site from the insert when constructing the biobrick to reduce the amount of base pairs in the primers. The primers also add 3 extra protective basepairs to the PstI site in the insert.


The construction of the final biobrick was initiated by cutting BBa_J04450 with XbaI, PstI, KnpI and FastAP and the insert with XbaI and PstI.

Source

Sequence adapted from Deinococcus Radiodurans. Ordered as synthetic gene.

References