Part:BBa_K3866004
pta- Phosphate acetyltransferase GB compatible with B3-B5
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 don't degrade 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.
Verification of Cloning
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1028
Illegal PstI site found at 1574
Illegal PstI site found at 1874
Illegal PstI site found at 2039 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1028
Illegal PstI site found at 1574
Illegal PstI site found at 1874
Illegal PstI site found at 2039 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1678
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1028
Illegal PstI site found at 1574
Illegal PstI site found at 1874
Illegal PstI site found at 2039 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1028
Illegal PstI site found at 1574
Illegal PstI site found at 1874
Illegal PstI site found at 2039
Illegal AgeI site found at 134
Illegal AgeI site found at 532
Illegal AgeI site found at 1778
Illegal AgeI site found at 1976 - 1000COMPATIBLE WITH RFC[1000]
Source
Synthesized by Twist Biosciences.
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).
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