Coding

Part:BBa_K1616013

Designed by: Johanna Chesnel   Group: iGEM15_IONIS_Paris   (2015-09-18)

ccdB reversed - RBS and double T7 terminator

ccdB

ccdB comes from the ccd system in Escherichia coli F plasmid and acts as a gyrase poison. The CcdB protein, constitutively expressed by P1010, is lethal to most of the BioBrick cell strains, only DB3.1 is resistant.


Reverse sequence


This part have created in order to make an efficient kill-switch. Our kill switch was designed to work in a system like pDawn, a light-inducible expression system. However, this system works with reversed sequence. That is why we designed a simple reversed Biobrick ccdB.
This composite part is composed of the reverse sequence of ccdB with reverse RBS (BBa_K1616011) and the double terminator T7 reversed (BBa_B0025). This part has been created in order to work with a reversed promoter, which will allow the synthesized of ccdB toxin.


Fig. 1: ccdB reversed


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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 226


Design Notes

All BioBrick parts used for assembling the composite part are compatible with the RFC10 Biobrick standard.


Source

This part is composed of ccdB reversed, RBS reversed and the double terminator T7 reversed.

[edit]
Categories
//biosafety/kill_switch
Parameters
None