Difference between revisions of "Part:BBa K3505021"

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[[File:T--Thessaly--djla.png|900px|thumb|none|<i><b>Fig.1:</b>DjlA</i>]]
 
[[File:T--Thessaly--djla.png|900px|thumb|none|<i><b>Fig.1:</b>DjlA</i>]]
  
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[[File:T--Thessaly--GB-CCAT-GCTT.jpeg|800px|thumb|none|<i><b>Fig.2:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
 
  
  
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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 pUPD2,<bbpart>BBa_K3505007</bbpart> 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 Golden Braid cloning.  
 
The CDS has position B2-B5.
 
The CDS has position B2-B5.
 +
[[File:T--Thessaly--GB-CCAT-GCTT.jpeg|800px|thumb|none|<i><b>Fig.2:</b>The overhangs of this part in the GoldenBraid Grammar</i>]]
  
 
===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 DjlA U4 C4 Digested with EcoRI, PstI Expected bands  2029, 894</i>]]
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[[File:T--Thessaly--d0.png|600px|thumb|none|<i><b>Fig.3:</b>Level 0 DjlA U4 C4 Digested with EcoRI, PstI Expected bands  2029, 894</i>]]
  
 
===Experimental Use and Experience===
 
===Experimental Use and Experience===
  
 
This part is used in <bbpart>BBa_K3505039</bbpart>
 
This part is used in <bbpart>BBa_K3505039</bbpart>
 +
 +
===Source===
 +
Georgios Skretas [1]
  
 
===Sequence and Features===
 
===Sequence and Features===

Latest revision as of 00:12, 28 October 2020


DjlA- DnaJ-like protein A GB compatible with B2-B5

Level 0 CDS

Fig.1:DjlA


Usage and Biology

DnaJ-like protein A (DjiA) is a membrane-bound DnaK co-chaperone that has antitoxic capacities for big protein expression in bacteria.[1]


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 Golden Braid 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 DjlA U4 C4 Digested with EcoRI, PstI Expected bands 2029, 894

Experimental Use and Experience

This part is used in BBa_K3505039

Source

Georgios Skretas [1]

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]


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.