Difference between revisions of "Part:BBa K4429005:Design"

 
 
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===Design Notes===
 
===Design Notes===
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-
 
  
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Ensuring the correct V->C region interface. Used Uniprot entry for human IGHG1 to find where constant region starts.
 +
 +
Removing BioBrick illegal restriction sites using silent mutation in SnapGene.
 +
 +
Mutation of Val214 to a GTT [2]
 +
 +
Addition of 6x polyhistidene tag at C-terminus.
  
  
 
===Source===
 
===Source===
  
 +
Variable region protein sequence from PDB 5AAW [1], reverse translated using E. coli codon usage table in SnapGene, constant region from GenBank, accession no: MT470197.1. The entire heavy chain was then codon optimised for E. coli K12 using IDT codon optimisation tool.
 
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-
  
 
===References===
 
===References===
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 +
[1] https://www.rcsb.org/structure/5AAW
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 +
[2] Leith, E. M., O'Dell, W. B., Ke, N., McClung, C., Berkmen, M., Bergonzo, C., Brinson, R. G., & Kelman, Z. (2019). Characterization of the internal translation initiation region in monoclonal antibodies expressed in Escherichia coli. The Journal of biological chemistry, 294(48), 18046–18056. https://doi.org/10.1074/jbc.RA119.011008

Latest revision as of 03:11, 14 October 2022


Ab513 IgG1 HC


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

Ensuring the correct V->C region interface. Used Uniprot entry for human IGHG1 to find where constant region starts.

Removing BioBrick illegal restriction sites using silent mutation in SnapGene.

Mutation of Val214 to a GTT [2]

Addition of 6x polyhistidene tag at C-terminus.


Source

Variable region protein sequence from PDB 5AAW [1], reverse translated using E. coli codon usage table in SnapGene, constant region from GenBank, accession no: MT470197.1. The entire heavy chain was then codon optimised for E. coli K12 using IDT codon optimisation tool. -

References

[1] https://www.rcsb.org/structure/5AAW

[2] Leith, E. M., O'Dell, W. B., Ke, N., McClung, C., Berkmen, M., Bergonzo, C., Brinson, R. G., & Kelman, Z. (2019). Characterization of the internal translation initiation region in monoclonal antibodies expressed in Escherichia coli. The Journal of biological chemistry, 294(48), 18046–18056. https://doi.org/10.1074/jbc.RA119.011008