Part:BBa_K1298004:Design
PgeChia 1-1 (insert) using J04500
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 296
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 822
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 682
Illegal NgoMIV site found at 851 - 1000INCOMPATIBLE 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.