Difference between revisions of "Part:BBa J45002:Design"
 
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===Design Notes===
 
===Design Notes===
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*There are two Met codons at the N-terminus of the protein (putative codons 1 and 4).  It is believed that the first Met codon is the actual start of the protein.  Reference:
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*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 <cite>Murfitt</cite>
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<biblio>
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#Dudareva pmid=11051108
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−
</biblio>
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−
#Forward primer (5' -> 3'): GTT TCT TCG AAT TCG CGG CCG CTT CTA G<b>AT GAA AGT GAT GAA GAA AC </b>
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#Reverse primer (5' -> 3'): GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TA<b>T CTC CTA CTT AGA GAA ACT AC </b>
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−
 
+
−
 
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*Bases 1-18 of the original ''A. majus'' sequence (gccggacgccaaagaaaa) were deleted.  Thus, this sequence begins with the first start codon.
 
*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.
 
*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
 
*The original stop codon (TGA at bp 1093) was deleted and a double TAA stop was inserted
 +
*Constructed by PCR
  
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===Source===
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====Template====
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Species: ''Antirrhinum majus'' (Snapdragon flower)
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We received a plasmid encoding the BAMT cDNA from Natalia Dudareva, Department of Horticulture and Landscape Architecture, Purdue University.
  
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Genbank accession number(s): [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=9789276 AF198492]
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====Forward primer====
 +
<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAA AGT GAT GAA GAA AC''' -3'</code>
  
−
We got this cDNA from Natalia Dudareva, Department of Horticulture and Landscape Architecture, Purdue University.
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====Reverse primer====
 +
<code>5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TA'''T CTC CTA CTT AGA GAA ACT AC''' -3'</code>
 +
 
 +
===Source===
 +
''BAMT'' gene from ''Antirrhinum majus'' (snapdragon flower) ([http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=9789276 GenBank AF198492])
  
 
===References===
 
===References===

Latest revision as of 21:33, 9 March 2008

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>