Difference between revisions of "Part:BBa K3866004"

 
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===Usage and Biology===
 
===Usage and Biology===
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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.
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The lacI has the ability to bind to the lac operator (<bbpart>BBa_R0010</bbpart> and <bbpart>BBa_R0011</bbpart> hybrid regulator) inhibiting the expression of the desirable transcription unit.
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===Design Notes===
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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.
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The CDS has position B3-B5.
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<p style="text-align: center;">
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    [[Image: T--Thessaly--GB-AATG-GCTT.jpeg|900px|thumb|none|<i><b>Figure 1.</b>The overhangs of this part in the GoldenBraid Grammar.</i>]]
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===Verification of Cloning===
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[[File:T--Thessaly--ptagel.png|700px|thumb|none|<i><b>Fig.3:</b>(U=Uncut C=Cut) Positive clone. 4 Expected bands 3027,1174, 50</i>]]
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===Sequence and Features===
  
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<span class='h3bb'>Sequence and Features</span>
 
 
<partinfo>BBa_K3866004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3866004 SequenceAndFeatures</partinfo>
  
  
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===Source===
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===Functional Parameters===
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<partinfo>BBa_K3866004 parameters</partinfo>
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Synthesized by Twist Biosciences.
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===References===
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*Lewis M. The lac repressor. <em>C R Biol.</em> 2005 Jun;328(6):521-48. doi: 10.1016/j.crvi.2005.04.004. PMID: 15950160.
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*Elowitz, M.B., Leibler, S. A synthetic oscillatory network of transcriptional      regulators. <em>Nature</em> 403, 335-338 (2000).

Revision as of 13:57, 25 September 2021


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.


Figure 1.The overhangs of this part in the GoldenBraid Grammar.


Verification of Cloning

Fig.3:(U=Uncut C=Cut) Positive clone. 4 Expected bands 3027,1174, 50

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1678
  • 23
    INCOMPATIBLE 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
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE 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).