Difference between revisions of "Part:BBa J45004:Design"
(→‎Template)
 
(11 intermediate revisions by 2 users not shown)
Line 6: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
Just CDS taken from GenBank
+
Constructed by PCR.
  
−
Designed Forward Strand (1 and 2):
+
====Template====
−
5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G <b>ATG GAA GTT GTT GAA GTT C</b> -3'<br>
+
pET-28a-''BSMT1''<cite>Negre03</cite>
−
*LENGTH: 19 <br>
+
*We thank Natalia Dudareva, Department of Horticulture and Landscape Architecture, Purdue University for the gift of the expression vector with the petunia ''BSMT1'' coding sequence.
−
*GC CONTENT: 36.8 % <br>
+
−
*MELT TEMP: 47.8 ºC <br>
+
−
<br>
+
−
Designed Reverse Strand (phBSMT1): 5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TA<b>T TAA TTT ATT TTG GTC AAG GAG</b> -3'<br>
+
−
*LENGTH: 22 <Br>
+
−
*GC CONTENT: 27.3 % <br>
+
−
*MELT TEMP: 46.6 ºC <br>
+
−
<br>
+
  
−
===Source===
+
====Forward primer====
 +
<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GGA AGT TGT TGA AGT TC''' -3'</code>
  
−
Species: Petunia x hybrida from GenBank (AY233465); (also available: A. thaliana and N. suaveolens)
+
====Reverse primer====
 +
<code>5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TA'''T TAA TTT ATT TTG GTC AAG GAG''' -3'</code>
 +
 
 +
===Source===
 +
*''BSMT1'' gene from ''Petunia x hybrida'' (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&val=28629494 AY233465])
  
 
===References===
 
===References===

Latest revision as of 22:29, 7 March 2008

SAM:benzoic acid/salicylic acid carboxyl methyltransferase I; converts salicylic acid to methyl sali


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 663
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 204


Design Notes

Constructed by PCR.

Template

pET-28a-BSMT1Negre03

  • We thank Natalia Dudareva, Department of Horticulture and Landscape Architecture, Purdue University for the gift of the expression vector with the petunia BSMT1 coding sequence.

Forward primer

5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GAT GGA AGT TGT TGA AGT TC -3'

Reverse primer

5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TAA TTT ATT TTG GTC AAG GAG -3'

Source

  • BSMT1 gene from Petunia x hybrida (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&val=28629494 AY233465])

References

<biblio>

  1. Underwood05 pmid=15849311
  2. Negre03 pmid=14630969

</biblio>