Coding

Part:BBa_K2244005:Design

Designed by: Chen Hong   Group: iGEM17_SSTi-SZGD   (2017-10-22)
Revision as of 07:19, 27 October 2017 by Qiucheng (Talk | contribs)

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LEV1 repressor


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 677
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

When designed the gene sequence of this fusion protein, the entire sequence of VVD, containnig a light-oxygen-voltage (LOV) domain & N cap, was added to the N-terminal of DNA-binding domain of LexA repressor. The C-terminal domain of lex repressor was removed.


Source

LEV1 repressor is a fusion protein of VVD and LexA, LexA repressor is a transcriptional repressor of SOS regulon in E.coli. It’s form chromosome of Escherichia coli str. K-12 substr. MG1655 (strain: K-12, substrain: MG1655) LexA autocleavage, stimulated by RecA, of the first 84 aa of LexA removes the DNA binding region and is required to activate the SOS response. LexA is a protein that belongs to the LexA family .

Blue light sensor VIVID was derived from the chromosome of Neurospora crassa. The LOV domain of the protein VVD has the capacity to self-dimerize upon light stimulation, Based on this property, the VVD LOV domain was fused with a smaller version of the Gal4 DNA binding domain and the p65 transactivation domain. A common feature of several blue light photoreceptors is the presence of LOV domains, which are able to bind a molecule of flavin mononucleotide (FMN) or flavin adenine dinucleotide (FAD) as chromophore, forming upon light stimulation a cysteinyl flavin C4a adduct.