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An esabox is an 18bp sequence to which the transcriptional regulator EsaR can bind. We placed one esabox right after the normally constitutive Anderson promoter [Bba_J23118] to create an EsaR repressible promoter. Transcription can be initiated by the specific AHL EsaR responds to [N-(3-oxo-hexanoyl)-L-homoserine lactone].

When characterising this part, we used the D91G variant of EsaR, which has been documented to be more responsive to lower AHL concentrations compared to the wild type EsaR [1]. This EsaR variant was obtained through addgene, and can be found on the registry [BBa_K2116001].

Two copies of EsaR were placed under the control of individual constitutive promoters [Bba_J23118] and expressed on a medium-low copy plasmid (replication origin pBR322/rop). Based on the 2013 paper by Shong&Collins we decided to test AHL concentrations in the range of 0 to 10 000nM, since in all combinations of esabox placements tested in this paper showed saturation at 10 000nM as seen below:

Dose response curves of EsaR, tested on promoters with various esabox placements. (Shong&Collins 2013 [1]).

We could not observe the same saturation behaviour in our tests,

Dose response curve of EsaR.

References: [1] Shong, Jasmine and Cynthia H. Collins. "Engineering The Esar Promoter For Tunable Quorum Sensing-Dependent Gene Expression". ACS Synth. Biol. 2.10 (2013): 568-575.

Sequence and Features Status: 500 Content-type: text/html

Software error:

Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84.
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84.
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 85.
Compilation failed in require at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12.
BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12.
Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9.
BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9.
Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 8.
BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 8.
Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12.
BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12.
Compilation failed in require at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.
BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.

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