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Coding

Part:BBa_K1590002:Design

Designed by: Manuel Blank   Group: iGEM15_Dundee   (2015-09-08)


Human Haptoglobin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1114
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Choosing the correct codon-bias, remove 'illegal' restriction sites, ensure that (stop)codons are in frame, addition of standard prefix and suffix.

Primers for cloning Haptoglobin into overexpression vector pQE80-L. 

  Forward: 5' GCGC GGATCC TCTGCGCTGGGTGCGGTT 3'

  Reverse: 5' GCGC GGTACC TTAGTTTTCCGCGATGGT 3'

Primers for cloning Haptoglobin into vector for bacterial 2-hybrid system, pUT18. 

  Forward: 5' GCGC GGATCC ATGTCTGCGCTGGGTGCG 3'

  Reverse: 5' GCGC GAATTC GAGTTTTCCGCGATGGTTTTC 3'

Primers for cloning Haptoglobin into vector for bacterial 2-hybrid system, pT25. 

  Forward: 5' GCGC GGTACC TTAGTTTTCCGCGATGGT 3'

  Reverse: 5' GCGC GGATCC ATGAACCTGCTGTCTCCGATCCTGTCT 3'

Part was synthesised with appropriate prefix/suffix sequences, hence no separate primers were required for cloning it into pSB1C3.


Source

In silico designed and optimized version of the human gene product for production in E. coli strains. Synthetic gene synthesised thanks to IDT sponsorship of iGEM 2015.

References

Smith A, McCulloh RJ. (2015) Hemopexin and haptoglobin: allies against heme toxicity from hemoglobin not contenders. Front Physiol 6:187. doi: 10.3389/fphys.2015.00187.

Andersen CB, Torvund-Jensen M, Nielsen MJ, de Oliveira CL, Hersleth HP, Andersen NH, Pedersen JS, Andersen GR, Moestrup SK. (2012) Structure of the haptoglobin-haemoglobin complex. Nature 489(7416):456-9. doi:10.1038/nature11369.