Difference between revisions of "Part:BBa J45014:Design"
Line 6: Line 6:
  
 
===Design Notes===
 
===Design Notes===
 +
*Constructed by PCR.
 +
*Eliminated EcoRI site at base pair 412 by changing base pair 414 from A to G
 
*<partinfo>BBa_J45014</partinfo> differs from <partinfo>BBa_J45006</partinfo> by a single point mutation at base pair 1148 (T -> C).
 
*<partinfo>BBa_J45014</partinfo> differs from <partinfo>BBa_J45006</partinfo> by a single point mutation at base pair 1148 (T -> C).
  
−
====New forward primer====
+
====Template====
−
<code>
+
''Saccharomyces cerevisiae'' strain ACLY387 genome
−
5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAA TGA AAT CGA TGA GAA AAA TC''' -3'
+
−
</code>
+
  
−
====New reverse primer====
+
====Forward primer====
−
<code>
+
<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAA TGA AAT CGA TGA GAA AAA TC''' -3'</code>
−
5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT '''TAa ggg cct aaa agg aga gct ttg''' -3'
+
 
−
</code>
+
====Reverse primer====
 +
<code>5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT '''TAa ggg cct aaa agg aga gct ttg''' -3'</code>
 +
 
 +
====Forward mutagenesis primer====
 +
<code>5'- GAA GCA AAT ATT AGA AGA GTT CAA AAA TAG TAA GGG -3'</code>
 +
 
 +
====Reverse mutagenesis primer====
 +
<code>5'- CCC TTA CTA TTT TTG AAC TCT TCT AAT ATT TGC TTC -3'</code>
  
 
===Source===
 
===Source===
−
''ATF1'' gene in ''Saccharomyces cerevisiae''.
+
''ATF1'' gene in ''Saccharomyces cerevisiae'' (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=Nucleotide&dopt=GenBank&val=1420813 Z75285])
  
 
===References===  
 
===References===  

Revision as of 22:03, 7 March 2008

alcohol acetyltransferase I; converts isoamyl alcohol to isoamyl acetate (banana odor)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 99
    Illegal BamHI site found at 1426
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 526
    Illegal SapI.rc site found at 1534


Design Notes

  • Constructed by PCR.
  • Eliminated EcoRI site at base pair 412 by changing base pair 414 from A to G
  • BBa_J45014 differs from BBa_J45006 by a single point mutation at base pair 1148 (T -> C).

Template

Saccharomyces cerevisiae strain ACLY387 genome

Forward primer

5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GAT GAA TGA AAT CGA TGA GAA AAA TC -3'

Reverse primer

5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TAa ggg cct aaa agg aga gct ttg -3'

Forward mutagenesis primer

5'- GAA GCA AAT ATT AGA AGA GTT CAA AAA TAG TAA GGG -3'

Reverse mutagenesis primer

5'- CCC TTA CTA TTT TTG AAC TCT TCT AAT ATT TGC TTC -3'

Source

ATF1 gene in Saccharomyces cerevisiae (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=Nucleotide&dopt=GenBank&val=1420813 Z75285])

References

<biblio>

  1. Horton03 pmid=12937998
  2. Singh--2007 pmid=17891501

</biblio>