Part:BBa_J45002:Design

SAM:benzoic acid carboxyl methyltransferase; converts benzoic acid to methyl benzoate (floral odor)

Assembly Compatibility:

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

  Illegal NheI site found at 970
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 654
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

• There are two methionine codons at the N-terminus of the protein (putative codons 1 and 4). It is believed that the first methionine codon is the actual start of the protein Murfitt
• Bases 1-18 of the original A. majus sequence (gccggacgccaaagaaaa) were deleted. Thus, this sequence begins with the first start codon.
• A silent mutation (A -> G) was made at base 702 to eliminate an SpeI cut site.
• The original stop codon (TGA at bp 1093) was deleted and a double TAA stop was inserted
• Constructed by PCR

Template

We received a plasmid encoding the BAMT cDNA from Natalia Dudareva, Department of Horticulture and Landscape Architecture, Purdue University.

Forward primer

5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GAT GAA AGT GAT GAA GAA AC -3'

Reverse primer

5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TAT CTC CTA CTT AGA GAA ACT AC -3'

Source

BAMT gene from Antirrhinum majus (snapdragon flower) ([http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=9789276 GenBank AF198492])

References

<biblio>

1. Murfitt pmid=11051108
2. Dudareva pmid=10852939

</biblio>