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Revision as of 19:33, 9 October 2020

LacI-Lac repressor GB compatible with B3-B5

Coding region for the LacI protein without an RBS.

Usage and Biology

The lac inhibitor and its operator is usually use for gene regulation and together they form a genetic circuit which called : the lac operon. In bacteria ,in abundance of lactose , lactose binds to lacI and the lac operon is activated. In synthetic biology , as an inducer we use IPTG, which doesn’t degraded in the cell , instead of lactose . IPTG is a molecular mimic of a lactose metabolite, which activates transcription of the lac operon. The lacI has the ability to bind to the lac operator (BBa_R0010 and BBa_R0011 hybrid regulator) inhibiting the expression of the desirable transcription unit.

Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is presents in pUPD2 and has overhangs compatible for Golden Braid cloning. The CDS has position B3-B5.


T--Thessaly--GBcloninggrammar.jpeg
The pre-existing part BBa_C0012has a C-terminal peptide that mediates degradation in bacteria (LVA tail). Our sequence doesn't contain LVA tail.

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
    COMPATIBLE WITH RFC[1000]


Source

E.Coli , sequence found through Addgene https://www.addgene.org/82021/

References

  • Lewis M. The lac repressor. C R Biol. 2005 Jun;328(6):521-48. doi: 10.1016/j.crvi.2005.04.004. PMID: 15950160.
  • Elowitz, M.B., Leibler, S. A synthetic oscillatory network of transcriptional regulators. Nature 403, 335-338 (2000).