Part:BBa_J45006:Design

alcohol acetyltransferase I (ATF1)

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

• Eliminated EcoRI site at base pair 412 by changing base pair 414 from A to G
• forward primer: 5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GAT GAA TGA AAT CGA TGA G -3'
• reverse primer: 5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TAA GGG CCT AAA AGG -3'

WE WANT TO REORDER: new forward and new reverse primers (6/26)

• new forward primer: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA GAT GAA TGA AAT CGA TGA GAA AAA TC -3'
• new 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

Source: S. cerevisiae, chromosome XV

Sequence obtained from Genbank, accession number [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>