Intermediate

Part:BBa_K1976051

Designed by: Alana Gouveia, Franziska Hameister, Jonas Sindlinger, Maik Schork   Group: iGEM16_TU_Darmstadt   (2016-10-14)

Colicin E2 DNase Trypsin Fragment with B0034 RBS

The DNase trypsin fragment (part BBa_K1976049) can be obtained by trypsin digestion of colicin E2. In this part the gene is fusioned to the strong ribosomal binding site B0034.

Usage

This part can be used for assembly of generators. In combination with the immunity protein Im2 (part BBa_K1976026) it serves as a safety mechanism (see part BBa_K1976053).

Characteristics

The DNase trypsin fragment of colicin E2 consists of 137 residues what conforms to about one third to colicin. We used only one of the three domains of colicin since the metabolic burdon would be significant smaller. This fragment differs to the part 50 in 5 additional amino acids since a trypsin digestion described in the literature leaded to this fragment. So we wanted to check out any difference in the activities between the DNase fragment with and without theese additional amino acids. The activity tests showed that the so called miniColicin has functional DNase activity.

Characteristics

Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 64
  • 1000
    COMPATIBLE WITH RFC[1000]


References
[1] Cascales et al, Colicin Biology, Microbiology and Molecular Biology Reviews, vol. 71, pp. 158-229, 2007

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