Difference between revisions of "Part:BBa K3505039"
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DjlA under control of arabinose inducible promoter. Having the ability to overexpress the CDS.
 
DjlA under control of arabinose inducible promoter. Having the ability to overexpress the CDS.
  
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[[File:T--Thessaly--d1map.png|600px|thumb|none|<i><b>Fig.1:</b>U1 C1 ParaBAD-DjlA-Terminator</i>]]
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[[File:T--Thessaly--paraBAD-DjIA.PHOT.png|600px|thumb|none|<i><b>Fig.1:</b>U1 C1 ParaBAD-DjlA-Terminator</i>]]
  
 
===Usage and Biology===
 
===Usage and Biology===
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===Design Notes===
 
===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 present in <bbpart>BBa_K3505009</bbpart> and has overhangs compatible for Golden Braid cloning.  
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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 cloned in <bbpart>BBa_K3505009</bbpart> and has overhangs compatible for Golden Braid cloning.  
  
  

Revision as of 11:15, 26 October 2020


ParaBAD-DjlA-Terminator

DjlA under control of arabinose inducible promoter. Having the ability to overexpress the CDS.

Fig.1:U1 C1 ParaBAD-DjlA-Terminator

Usage and Biology

This Trancriscription Unit can overexpress antitoxic proteins in order to to achieve epression of non omologous membrane proteins in E. coli.


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 cloned in BBa_K3505009 and has overhangs compatible for Golden Braid cloning.



Verification of cloning

Fig.2:U1 C1 ParaBAD-DjlA-Terminator Digested with EcorV, BamHI. Expected bands 4212, 853

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1148
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 983
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 965


References

  • [1]Gialama, D., Delivoria, D., Michou, M., Giannakopoulou, A. and Skretas, G., 2017. Functional Requirements for DjlA- and RraA-Mediated Enhancement of Recombinant Membrane Protein Production in the Engineered Escherichia coli Strains SuptoxD and SuptoxR. Journal of Molecular Biology, 429(12), pp.1800-1816.