Difference between revisions of "Part:BBa K3505022"
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[[File:T--Thessaly--rraa.png|900px|thumb|none|<i><b>Fig.1:</b>RraA</i>]]
 
[[File:T--Thessaly--rraa.png|900px|thumb|none|<i><b>Fig.1:</b>RraA</i>]]
  
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[[File:T--Thessaly--GB-CCAT-GCTT.jpeg|700px|thumb|none|<i><b>Fig.2:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
 
  
  
 
===Usage and Biology===
 
===Usage and Biology===
−
Regulator of ribonuclease activity A (RraA), an repressor of the mRNA-degrading ability of the E. coli RNase E.[1] This
+
The Regulator of ribonuclease activity A (RraA), a repressor of the mRNA-degrading ability of the <i>E. coli</i> RNase E. [1] This
−
protein leads to better expression of non omologous membrane proteins in E. coli.
+
protein leads to better expression of non omologous membrane proteins in <i>E. coli</i>.
 +
 
 
===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 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 cloned in pUPD2 <bbpart>BBa_K3505007</bbpart> and has overhangs compatible for GoldenBraid cloning.  
 
The CDS has position B2-B5.
 
The CDS has position B2-B5.
−
 
+
[[File:T--Thessaly--GB-CCAT-GCTT.jpeg|700px|thumb|none|<i><b>Fig.2:</b>The overhangs of this part in the GoldenBraid Grammar</i>]]
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+
  
 
===Verification of cloning===
 
===Verification of cloning===
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[[File:T--Thessaly--d0.png|600px|thumb|none|<i><b>Fig.4:</b>Level 0 RraA U2 C2 Digested with EcoRI, PstI Expected bands 2029, 564</i>]]
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[[File:T--Thessaly--d0.png|600px|thumb|none|<i><b>Fig.3:</b>Level 0 RraA U2 C2 Digested with EcoRI, PstI Expected bands 2029, 564</i>]]
  
 
===Experimental Use and Experience===
 
===Experimental Use and Experience===
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This part is used in <bbpart>BBa_K3505038</bbpart>
 
This part is used in <bbpart>BBa_K3505038</bbpart>
  
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====Sequence and Features===
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===Sequence and Features===
 
<partinfo>BBa_K3505022 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3505022 SequenceAndFeatures</partinfo>
 +
===Source===
 +
Georgios Skretas [1]
  
 
===References===
 
===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. <em>Journal of Molecular Biology</em>, 429(12), pp.1800-1816.
 
*[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. <em>Journal of Molecular Biology</em>, 429(12), pp.1800-1816.

Latest revision as of 00:14, 28 October 2020


RraA- Regulator of ribonuclease activity A GB compatible with B2-B5

Level 0 CDS

Fig.1:RraA


Usage and Biology

The Regulator of ribonuclease activity A (RraA), a repressor of the mRNA-degrading ability of the E. coli RNase E. [1] This protein leads to better expression 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 pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. The CDS has position B2-B5.

Fig.2:The overhangs of this part in the GoldenBraid Grammar

Verification of cloning

Fig.3:Level 0 RraA U2 C2 Digested with EcoRI, PstI Expected bands 2029, 564

Experimental Use and Experience

This part is used in BBa_K3505038

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

Georgios Skretas [1]

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.