Protein_Domain

Part:BBa_K392008:Experience

Designed by: Damian Barnard, Teoh Shao Thing, Tadashi Nakamura   Group: iGEM10_Osaka   (2010-10-24)

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K392008

User Reviews

UNIQ64e838201ce49ea0-partinfo-00000000-QINU

BBa_K392008 Allan Crossman (Edinburgh 2011)

This part is known to work in the Edinburgh lab and yet our copy has what appears to be an early frameshift. Of course, this is impossible. We therefore believe that the 2nd ATG present in the sequence (about 220 bases in) is the true start codon. There is a plausible ribosome binding site just upstream of it.

In support of this conclusion, the most recent [http://www.ncbi.nlm.nih.gov/nuccore/332337569?from=3105074&to=3106528 published sequence] of the gene starts at that location. It also incorporates a total of three extra bases at various locations, which mean that the middle of the protein sequence has very different amino acids from the official sequence of K392008 (until it comes back into the same frame).

Although the centre of the part is not readily accessible to Sanger sequencing using the standard sequencing primers, we can at least confirm the presence of one of these bases. It is likely that the true sequence (i.e. present in physical DNA) of K392008 is similar to the published NCBI sequence in the link above (but with the aforementioned ~220 additional upstream bases as well).

We created BBa_K523010 based on this information.

;

UNIQ64e838201ce49ea0-partinfo-00000003-QINU