Difference between revisions of "Part:BBa J45001:Design"

(→‎Design Notes)
 
(4 intermediate revisions by 3 users not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J45001 short</partinfo>
 
<partinfo>BBa_J45001 short</partinfo>
Line 7: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
#no restriction enzyme cut sites in coding region
+
*No BioBrick restriction enzyme cut sites in coding region
−
#changed natural stop codon to TAA TAA
+
*Changed natural stop codon to TAA TAA
−
#primer design
+
*Constructed by PCR
−
#*forward = 5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G<b>AT GAC AAA ACA AAC ACA AAA G</b> -3'
+
 
−
#*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'
+
====Forward primer====
−
#*internal sequencing primer = 5' - gga atg ctg cta tat ttg gg
+
<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAC AAA ACA AAC ACA AAA G''' -3'</code>
 +
 
 +
====Reverse primer====
 +
<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>
 +
 
 +
====Internal sequencing primer====
 +
<code>5'- gga atg ctg cta tat ttg gg -3'</code>
  
 
===Source===
 
===Source===
−
#GenBank Accession No. AF515284
+
''SAMT'' gene from ''Antirrhinum majus'' (snapdragon flower) ([http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=23957317 GenBank AF515284])
−
#Species = Antirrhinum Majus, Snapdragon
+
  
 
===References===
 
===References===
−
#corresponding author: Natalia Dudareva
+
<biblio>
−
#Pubmed: PMID: 12361714
+
#Negre-ArchBiochemBiophys-2002 pmid=12361714
−
#Abstract for SAMT enzyme:
+
</biblio>
−
##http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12361714&dopt=Abstract
+
 
−
#Wiki Link:
+
*[http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-6-6 MIT iGEM 2006 notebook entry]
−
##http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-6-6
+

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]