Part:BBa_K4477011:Design

Human/mouse anti-apoB(MDA) (anti-oxLDL) IgG - complete expression cassette

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 826
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1911
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 455
  

Design Notes

Amino acid sequences reverse transcribed and codon optimized for expression in E. coli B (the parent strain of SHuffle). Illegal restriction enzymes were removed via codon optimization.

A two-way terminator was used so that any genes in the vector in which this insert will be inserted that happen to be on the strand opposite to the strand encoding the insert will not be read backward into the insert and interfere with transcription of our antibody of interest.

For detailed annotations, see Team Virginia 2022's Benchling folder here: https://benchling.com/mrkhoury/f_/YR4zZSUb-virginia-igem-2022-dna-constructs/

Source

Orticumab light chain amino acid sequence acquired from IMGT: https://www.imgt.org/3Dstructure-DB/cgi/details.cgi?pdbcode=9635
Orticumab heavy chain amino acid sequence acquired from IMGT: https://www.imgt.org/3Dstructure-DB/cgi/details.cgi?pdbcode=9635
Mouse heavy chain amino acid sequence acquired from NCBI: Gene ID: 16017

References

1. 10.1038/ncomms9072
2. https://www.clinicaltrials.gov/ct2/show/NCT04776629
3. https://abcentra.com/our-science/