Part:BBa_K1590006:Design

Human Lanosterol Synthase

Assembly Compatibility:

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

  Illegal NheI site found at 819
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 666
  Illegal AgeI site found at 145
  Illegal AgeI site found at 283
  
• 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.

Intermediate primers for sequencing Lanosterol Synthase.

  Forward: GGCTGTTCCCGGACTGGG

  Reverse: CGGTGACATCATGATCGAC

Primers for cloning Lanosterol Synthase into overexpression vector pQE80-L.

  Forward: GCGCGGATCCATGACCGAAGGAACCTGC

  Reverse: GCGCGGTACCTTACGGGTGACCCGCCAGCGC

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

Source

In silico designed gene optimised for expression in an E. coli chassis. The sequence wa suppplied through the kind sponosrship of iGEM 2015 by IDT.

References

Thoma R, Schulz-Gasch T, D'Arcy B, Benz J, Aebi J, Dehmlow H, Hennig M, Stihle M, Ruf A. (2004) Insight into steroid scaffold formation from the structure of human oxidosqualene cyclase. Nature 432(7013):118-22.