Composite

Part:BBa_K1298004:Design

Designed by: CoBRA Team   Group: iGEM14_CoBRA   (2014-06-19)


PgeChia 1-1 (insert) using J04500


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 296
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 822
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 682
    Illegal NgoMIV site found at 851
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 996


Design Notes

DNA optimization was required to remove illegal cut sites. This was successful. There is also no secretory tag, meaning that the protein (chitinase) is not being transported out of the cell, and instead staying inside.


Source

BBa_K1298001 (PgeChia 1-1)

  • Phytochemistry paper of Dr. Kolosova (1)
  • NCBI (protein code) (2)
  • Bio Basic Inc. (DNA optimization)

References

1. Kolosova, N. “Cloning and characterization of chitinases from interior spruce and lodgepole pine.” 1 Oct. 2013. Feb. 2014 <http://www.ncbi.nlm.nih.gov/pubmed/24564978>

2. Kolosova, N. "Picea engelmannii x Picea glauca class I chitinase (Chia1-1) mRNA, complete cds." 30 Dec. 2013. Feb. 2014 <http://www.ncbi.nlm.nih.gov/nuccore/HM219843>*

*The sequence found at this link has been optimized to remove illegal cut sites (EcoRI, XbaI, SpeI, PstI) and therefore is not the same as the sequence found on this part page.