Generator

Part:BBa_K1976054

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

Colicin E2 DNase and T7 promoter

This part encodes the DNase fragment of colicin E2 with the RBS B0034 (part BBa_K1976048) and a T7 promoter.

Figure 1:Figure 5: growth of BL21 E. coli transformed with a): BBa_K1976054 and b): BBa_K1976055.


Usage

The colicin DNase fragment is an active DNase and can be used for DNA degradation. The T7 promoter is an inducable promoter wich ensures expression of the so called miniColicin after induction with IPTG in cells which have a T7 promoter system.

Characteristics

The DNase fragment of colicin E2 has a molecular weight of 15.2 kDa and consists of 133 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. The activity tests showed that the so called miniColicin has functional DNase activity.

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 66
  • 1000
    COMPATIBLE WITH RFC[1000]


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

[edit]
Categories
Parameters
None