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Coding

Part:BBa_K1590007:Design

Designed by: Conner Craigon   Group: iGEM15_Dundee   (2015-09-08)


Human Odorant Binding Protein 2A


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 445
  • 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 Odorant Binding Protein 2A into pSB1C3 and into overexpression vector pQE80-L. 

  Forward: GCGC GAATTCGCGGCCGCTTCTAG A TGCTGTCTTTCACCCTGGAAGAA
  Reverse: GCGC CTGCAGCGGCCGCTACTAGT ATTATTAGTGTTCCAGAACGCAAG

Part was synthesised as coding sequence only. Prefix, suffix, and stop codons were added by using the primers above.


Furthermore, another set of primers was used in order to subclone 2 parts of the sequence that lead to structurally distinct parts of resultant protein.

Primers for cloning Odorant Binding Protein 2A - Helix into vector for bacterial 2-hybrid system, pUT25. 

   Forward: GCGC GGATCC GCCGAACACCAACCTGGAAGCG
   Reverse: GCGC GGTACC TTAGTGTTCCAGAACGCAAGA

Primers for cloning Odorant Binding Protein 2A - Barrel into vector for bacterial 2-hybrid system, pT18. 

   Forward: GCGC GAATTCGCGGCCGCTTCTAG A TGCTGTCTTTCACCCTGGAAGAA
   Reverse: GCGC CTGCAGCGGCCGCTACTAGT ATTATT AGTGTTCCAGAACGCAAG

Source

In silico optimised version for E. coli K12 strains. Sequence ordered through IDT.

References