Difference between revisions of "Part:BBa J45014:Design"

(→‎References)
(→‎Source)
 
(5 intermediate revisions by the same user not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J45014 short</partinfo>
 
<partinfo>BBa_J45014 short</partinfo>
Line 7: Line 6:
  
 
===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).
  
−
===Source===
+
====Template====
 +
''Saccharomyces cerevisiae'' strain ACLY387 genome
  
−
This part comes from <partinfo>BBa_J45006</partinfo>.
+
====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===
 +
''ATF1'' gene from ''Saccharomyces cerevisiae'' (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=Nucleotide&dopt=GenBank&val=1420813 Z75285])
  
−
===References===
+
===References===  
−
The publication below reports the use of ATF1 in ''E. coli'' to produce isoamyl acetate from isoamyl alcohol.  Note that the publication date was after the 2006 MIT iGEM designed, constructed and tested <partinfo>BBa_J45014</partinfo>.
+
 
<biblio>
 
<biblio>
 +
#Fujii--1984 pmid=8085822
 +
#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>