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

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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J45001 short</partinfo>
 
<partinfo>BBa_J45001 short</partinfo>
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===Design Notes===
 
===Design Notes===
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#changed initial stop codon to TAA TAA
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*No BioBrick restriction enzyme cut sites in coding region
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*Changed natural stop codon to TAA TAA
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*Constructed by PCR
  
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#primer design
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====Forward primer====
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#*forward = 5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GAG <b>ATG ACA AAA CAA ACA CAA AAG</b> -3'
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<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAC AAA ACA AAC ACA AAA G''' -3'</code>
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#*reverse = 5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TA<b>T TAG TTT CTT TTT GCT TG</b> -3'
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====Reverse primer====
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<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>
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====Internal sequencing primer====
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<code>5'- gga atg ctg cta tat ttg gg -3'</code>
  
 
===Source===
 
===Source===
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#GenBank Accession No. AF515284
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''SAMT'' gene from ''Antirrhinum majus'' (snapdragon flower) ([http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=23957317 GenBank AF515284])
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#Organism = Antirrhinum Majus, Snapdragon
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===References===
 
===References===
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#corresponding author: Natalia Dudareva
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<biblio>
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#Pubmed: PMID: 12361714
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#Negre-ArchBiochemBiophys-2002 pmid=12361714
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#Abstract for SAMT enzyme:
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</biblio>
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##http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12361714&dopt=Abstract
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#Wiki Link:
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*[http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-6-6 MIT iGEM 2006 notebook entry]
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##http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-6-6
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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]