Difference between revisions of "Part:BBa J45002:Design"

(→‎Design Notes)
 
(5 intermediate revisions by 2 users not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J45002 short</partinfo>
 
<partinfo>BBa_J45002 short</partinfo>
Line 7: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
*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:
+
*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>
−
<biblio>
+
−
#Dudareva pmid=11051108
+
−
</biblio>
+
−
 
+
 
*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
  
−
===Source===
+
====Template====
−
Genbank accession number(s): AF198492
+
We received a plasmid encoding the BAMT cDNA from Natalia Dudareva, Department of Horticulture and Landscape Architecture, Purdue University.
  
−
We got this cDNA from N Dudareva at Purdue.
+
====Forward primer====
 +
<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAA AGT GAT GAA GAA AC''' -3'</code>
 +
 
 +
====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===
 +
<biblio>
 +
#Murfitt pmid=11051108
 +
#Dudareva pmid=10852939
 +
</biblio>

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>