Difference between revisions of "Part:BBa K3407004:Design"

 
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===Design Notes===
 
===Design Notes===
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Considerations here
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Fox-1 amino acid sequence retrieved from NCBI residues 109–208 following the Literature’s materials and methods. As an N-HisTag from pET-28(a) was used in materials and methods, its amino acid sequence was added in the N-Terminal of Fox-1. The Cry7Ca1 gene was codon-optimized for expression in Escherichia coli using the GenSmartTM Codon Optimization Tool. The forbidden restriction enzymes sites were the following: BioBrick forbidden restriction sites (EcoRI, XbaI, SpeI, PstI) and Type IIS forbidden restriction sites  (BsaI, SapI and Bpil). BglBrick prefix (nt 1525-1560) and suffix (nt 2277-2318) from pBbB7a (Addgene #35358) were added to the optimised sequence generated. They were designed to be cloned with Gibson Assembly.
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===Source===
 
===Source===
  
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Source this part
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Plasmids used were from BglBrick repository. pBbB7a-GFP was a gift from Jay Keasling (Addgene plasmid # 35358 ; http://n2t.net/addgene:35358 ; RRID:Addgene_35358) <html><a href="#1">[1]</a></html>.
  
 
===References===
 
===References===
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<a id="1" href="https://jbioleng.biomedcentral.com/articles/10.1186/1754-1611-5-12" target="_blank">
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BglBrick vectors and datasheets: A synthetic biology platform for gene expression. Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD. J Biol Eng. 2011 Sep 20;5:12. 10.1186/1754-1611-5-12 PubMed 21933410</a>
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Revision as of 16:24, 26 October 2020


Fox-1 RBD: a protein domain binding strongly and specifically to RNA sequence UGCAUGU.


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]


Design Notes

Fox-1 amino acid sequence retrieved from NCBI residues 109–208 following the Literature’s materials and methods. As an N-HisTag from pET-28(a) was used in materials and methods, its amino acid sequence was added in the N-Terminal of Fox-1. The Cry7Ca1 gene was codon-optimized for expression in Escherichia coli using the GenSmartTM Codon Optimization Tool. The forbidden restriction enzymes sites were the following: BioBrick forbidden restriction sites (EcoRI, XbaI, SpeI, PstI) and Type IIS forbidden restriction sites (BsaI, SapI and Bpil). BglBrick prefix (nt 1525-1560) and suffix (nt 2277-2318) from pBbB7a (Addgene #35358) were added to the optimised sequence generated. They were designed to be cloned with Gibson Assembly.


Source

Plasmids used were from BglBrick repository. pBbB7a-GFP was a gift from Jay Keasling (Addgene plasmid # 35358 ; http://n2t.net/addgene:35358 ; RRID:Addgene_35358) [1].

References

Ordered List

  1. BglBrick vectors and datasheets: A synthetic biology platform for gene expression. Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD. J Biol Eng. 2011 Sep 20;5:12. 10.1186/1754-1611-5-12 PubMed 21933410