Difference between revisions of "Part:BBa J45014:Design"
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===Design Notes===
 
===Design Notes===
−
*This part differs from ATF1 [https://parts.igem.org/wiki/index.php/Part:BBa_J45006 BBa_J45006] at one base pair, (1148; T -> C).
+
*Constructed by PCR.
−
*It is currently in the '''pSB1A3-1''' BioBrick backbone.
+
*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).
  
−
*new forward primer: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA G<b>AT GAA TGA AAT CGA TGA GAA AAA TC</b> -3'
+
====Template====
−
*new reverse primer: 5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT <b>TAa ggg cct aaa agg aga gct ttg</b> -3'
+
''Saccharomyces cerevisiae'' strain ACLY387 genome
 +
 
 +
====Forward primer====
 +
<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAA TGA AAT CGA TGA GAA AAA TC''' -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===
−
This part comes from <partinfo>BBa_J45006</partinfo>.
+
''ATF1'' gene from ''Saccharomyces cerevisiae'' (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=Nucleotide&dopt=GenBank&val=1420813 Z75285])
  
 
===References===  
 
===References===  
 
<biblio>
 
<biblio>
 +
#Fujii--1984 pmid=8085822
 
#Horton03 pmid=12937998
 
#Horton03 pmid=12937998
 
#Singh--2007 pmid=17891501
 
#Singh--2007 pmid=17891501
 
</biblio>
 
</biblio>

Latest revision as of 22:10, 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 from Saccharomyces cerevisiae (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=Nucleotide&dopt=GenBank&val=1420813 Z75285])

References

<biblio>

  1. Fujii--1984 pmid=8085822
  2. Horton03 pmid=12937998
  3. Singh--2007 pmid=17891501

</biblio>