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

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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J45014 short</partinfo>
 
<partinfo>BBa_J45014 short</partinfo>
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===Design Notes===
 
===Design Notes===
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*This part differs from ATF1 [https://parts.igem.org/wiki/index.php/Part:BBa_J45006 BBa_J45006] at one base pair, (1148; T -> C).
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*Constructed by PCR.
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*It is currently in the '''pSB1A3-1''' BioBrick backbone.
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*Eliminated EcoRI site at base pair 412 by changing base pair 414 from A to G
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*<partinfo>BBa_J45014</partinfo> differs from <partinfo>BBa_J45006</partinfo> by a single point mutation at base pair 1148 (T -> C).
  
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===Source===
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====Template====
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''Saccharomyces cerevisiae'' strain ACLY387 genome
  
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This part comes from <partinfo>BBa_J45006</partinfo>.
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====Forward primer====
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<code>5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''AT GAA TGA AAT CGA TGA GAA AAA TC''' -3'</code>
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====Reverse primer====
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<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====
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<code>5'- GAA GCA AAT ATT AGA AGA GTT CAA AAA TAG TAA GGG -3'</code>
 +
 
 +
====Reverse mutagenesis primer====
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<code>5'- CCC TTA CTA TTT TTG AAC TCT TCT AAT ATT TGC TTC -3'</code>
 +
 
 +
===Source===
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''ATF1'' gene from ''Saccharomyces cerevisiae'' (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=Nucleotide&dopt=GenBank&val=1420813 Z75285])
  
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===References===
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===References===  
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<biblio>
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#Fujii--1984 pmid=8085822
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#Horton03 pmid=12937998
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#Singh--2007 pmid=17891501
 +
</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>