Project

Part:BBa_K494003

Designed by: Haissi Cui, Tilman Flock, Sebastian Gude, Christoph Hartlmüller, Florian Praetorius, Jan Schüürmann, Tobi Wauer, Philipp Wortmann   Group: iGEM10_TU_Munich   (2010-10-25)

Exemplary insert for BBa_K494001 (His-Term)

In general we want to provide a new, scalable principle of gene regulation based on termination and antitermination which can be further developed, tested and optimizted by everybody. To enable that in the easiest way possible we decided to not just add the switches we designed but instead the material needed for evaluation. Therefore we focus on providing the parts needed for verification and testing of new individual switches.



This is a ready to use part consisting of our screening backbone BBa_K494001 and a His-terminator in front of and an IPTG inducible, corresponding signal is cloned following mCherry for testing reasons.
This part belongs to the screening system for evaluation of individual switches constructed on the principle of bioLOGICS, in this case BBa_K494004 may serve as a reference to compare results with those presented by the TU Munich 2010 iGEM Team. Together with BBaK494003 (Terminator and corresponding switch which leads to antitermination after IPTG induction) and BBa_K494002 (positive control for mCherry expression) it is a setup to evaluate termination and antitermination efficiency of the His-Terminator.


The His-Terminator (which does not exist as an individual BioBrick since there are enough thoroughly tested Terminators yet in the registry) is based on an attenuator stem loop in front of the his-operon of Salmonella enterica. It was found to be a strong terminator with allowed no detectable transcription of the following mCherry gene so this BioBrick can also be used as a reference for testing other PoPS devices.

Emission spectra of induced BBa_K494003 ; A: eGFP fluorescence ex: 501 nm, B: mCherry fluorescence ex: 587 nm

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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