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

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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J45002 short</partinfo>
 
<partinfo>BBa_J45002 short</partinfo>
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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 ATG AAA GTG ATG AAG AAA C
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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
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*Constructed by PCR
  
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===Source===
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====Template====
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Genbank accession number(s): AF198492
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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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We got this cDNA from N Dudareva at Purdue.
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====Forward primer====
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<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAA AGT GAT GAA GAA AC''' -3'</code>
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====Reverse primer====
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<code>5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TA'''T CTC CTA CTT AGA GAA ACT AC''' -3'</code>
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 +
===Source===
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''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
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#Dudareva pmid=10852939
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</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>