Difference between revisions of "Part:BBa K3505003"
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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 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 pre-existing part <bbpart>BBa_C0012</bbpart>has a C-terminal peptide that mediates degradation in bacteria (LVA tail). Our sequence doesn't contain LVA tail. | + | The pre-existing part<bbpart>BBa_C0012</bbpart>has a C-terminal peptide that mediates degradation in bacteria (LVA tail). Our sequence doesn't contain LVA tail. |
===Sequence and Features=== | ===Sequence and Features=== | ||
Revision as of 15:35, 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 pre-existing partBBa_C0012has a C-terminal peptide that mediates degradation in bacteria (LVA tail). Our sequence doesn't contain LVA tail.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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).