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Coding

Part:BBa_K1590001:Design

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


Human Haemoglobin B


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

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

Primers for cloning Haemoglobin A into overexpression vector pQE80-L. 

   Forward: 5' GCGC GGATCC GTTCACCTGACCCCGGAAG 3'
   Reverse: 5' GCGC GGTACC TTAGTGGTATTTGTGCGC 3'

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

   Forward: 5' GCGC GGATCC ATGGTTCACCTGACCCCGGAAG 3'
   Reverse: 5' GCGC GAATTC TATTAGTGGTATTTGTGCGCCAG 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 for E. coli expression. The synthetic gene was supplied thanks to generous sponsorship of iGEM 2015 by IDT.

References

Kidd RD, Russell JE, Watmough NJ, Baker EN, Brittain T. (2001) The role of beta chains in the control of the hemoglobin oxygen binding function: chimeric human/mouse proteins, structure, and function. Biochemistry 40(51):15669-75.

Muirhead H, Perutz MF. (1963) Structure of haemoglobin: a three-dimensional Fourier synthesis of reduced human haemoglobin at 5.5A resolution. Nature 199:633-8.