Coding

Part:BBa_K1659300:Design

Designed by: Wei Chung Kong   Group: iGEM15_Oxford   (2015-08-28)

Micrococcal Nuclease (DNase)


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 and Sources

DNase BLAST

BLAST search using translated sequence of BBa_K729004, from which only the nucleotide sequence that coded for the DNase was identified and isolated


The gene sequence for micrococcal DNase was obtained from BBa_K729004, a part comprising DsbA 2-19 fused to the N-terminal of micrococcal nuclease made by [http://2012.igem.org/Team:University_College_London Team UCL 2012]. The part sequence was not annotated, and as such we ran a [http://blast.ncbi.nlm.nih.gov/blast/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome Protein BLAST search] using the part's translated sequence to identify the bases that coded for our desired peptide [1]. The sequence for micrococcal DNase was adopted as-is from said part, without further codon optimization.

A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography.


References

[1] Stephen F. Altschul, Thomas L. Madden, Alejandro A. Schäffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman, 1997. "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25:3389-3402.