Arabinose inducible TorA-Reflectin A1-sfGFP generator
This construct is Reflectin A1 protein with a TorA signal peptide on the N terminus, and superfolder GFP (BBa_I746916) fused at the C-terminus. The reflectin A1 sequence is codon-optimized for E coli. The TorA signal peptide has been shown to allow export of GFP via the twin arginine pathway. The fusion gene is under the control of the pBAD promoter BBa_I0500, which is tightly controlled by two factors:
- L-arabinose monosaccharide taken up by the cell from the medium, which acts as an inducer.
- AraC protein included in the I0500 biobrick, which acts an a repressor.
Therefore, the araC-pBAD system offers regulatable control of gene expression in the presence of the inducer and highly repressed in the absence of the inducer. Read more about the pBAD and arabinose system in E.coli.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 1205
- 21Illegal BamHI site found at 1144
Illegal BamHI site found at 1369
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 979
- 1000COMPATIBLE WITH RFC
Usage and Biology
Cambridge 2011 were unable to test export fully as of the wiki freeze (21st Sep). See our wiki for more information on our experiments.
The protein coding sequence for reflectin originally came from cells of an edible squid. There have been no reported safety issues for reflectins, so we do not anticipate the need for extra precautions when using this BioBrick part. See our safety page for more information.
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