DNA

Part:BBa_K582000:Design

Designed by: Kettner Griswold, Mitesh Agrawal   Group: iGEM11_GeorgiaTech   (2011-09-28)

500bp of Spacer/Repeat Region of the CRISPR genome present in Strep. thermophilus LMD9 strain


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The entire Repeat/Spacer Region is 1400bp in the CRISPR sequence in the Streptococcus Thermophilus LMD-9 strain but it has a SpeI site at 500 bp. We failed in our attempt to conduct Site Directed Mutagenesis and modify the SpeI site. For this reason, for our insert, we cut the original 1400bp DNA at the 500bp and have decided to use that as our insert. We hypothesize that inserting the cas 9 genome on a separate shuttling vector along with the biobrick should result in the expression of the CRISPR system giving the bacteria resistance against the phages which the spacer sequences represent.

Source

http://www.ncbi.nlm.nih.gov/nuccore/116626972?report=genbank&from=648900&to=649423

This comes from the genomic sequence of Streptococcus Thermophilus LMD-9

References