Difference between revisions of "Part:BBa J45001:Design"
 
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===Design Notes===
 
===Design Notes===
 +
*No BioBrick restriction enzyme cut sites in coding region
 +
*Changed natural stop codon to TAA TAA
 +
*Constructed by PCR
  
−
<b> WE WANT TO REORDER: the reverse and the internal sequencing primers (6/26) </b>
+
====Forward primer====
 +
<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAC AAA ACA AAC ACA AAA G''' -3'</code>
  
−
#no restriction enzyme cut sites in coding region
+
====Reverse primer====
−
#changed natural stop codon to TAA TAA
+
<code>5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TA'''c tct cgc ctt gtc atg gaa ata g''' -3'</code>
−
#primer design
+
 
−
#*forward = 5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G<b>AT GAC AAA ACA AAC ACA AAA G</b> -3'
+
====Internal sequencing primer====
−
#*reverse = 5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TA<b>c tct cgc ctt gtc atg gaa ata g</b> -3'
+
<code>5'- gga atg ctg cta tat ttg gg -3'</code>
−
#*internal sequencing primer = 5' - gga atg ctg cta tat ttg gg
+
  
 
===Source===
 
===Source===
−
#Species: ''Antirrhinum majus'', Snapdragon flower
+
''SAMT'' gene from ''Antirrhinum majus'' (snapdragon flower) ([http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=23957317 GenBank AF515284])
−
#GenBank Accession: [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=23957317 AF515284]
+
  
 
===References===
 
===References===
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</biblio>
 
</biblio>
  
−
*Wiki Link http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-6-6
+
*[http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-6-6 MIT iGEM 2006 notebook entry]

Latest revision as of 21:41, 9 March 2008

SAM:salicylic acid carboxyl methyltransferase; converts salicylic acid to methyl salicylate (winter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 577
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 901
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 421
    Illegal NgoMIV site found at 425
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 471
    Illegal SapI.rc site found at 75


Design Notes

  • No BioBrick restriction enzyme cut sites in coding region
  • Changed natural stop codon to TAA TAA
  • Constructed by PCR

Forward primer

5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GAT GAC AAA ACA AAC ACA AAA G -3'

Reverse primer

5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TAc tct cgc ctt gtc atg gaa ata g -3'

Internal sequencing primer

5'- gga atg ctg cta tat ttg gg -3'

Source

SAMT gene from Antirrhinum majus (snapdragon flower) ([http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=23957317 GenBank AF515284])

References

<biblio>

  1. Negre-ArchBiochemBiophys-2002 pmid=12361714

</biblio>

  • [http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-6-6 MIT iGEM 2006 notebook entry]