Coding

Part:BBa_M36843:Design

Designed by: Jasmine, Katie, Matthew (Bluegreen)   Group: Stanford BIOE44 - S11   (2015-10-24)


E2F1 protein coding sequence


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1242
    Illegal PstI site found at 437
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 437
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 183
    Illegal BglII site found at 754
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1242
    Illegal PstI site found at 437
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1242
    Illegal PstI site found at 437
    Illegal AgeI site found at 145
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

There was a restriction site, Bbs1, presented in the original DNA sequence. Because of this, a single base pair in the sequence was changed from an A to a G. The codon still encodes for the same protein, Glutamic acid. The specific sequence was changed from GAAGAC to GAGGAC. Additionally, we used the website, https://www.idtdna.com/CodonOpt, to optimize the codons for S.cerevisiae. The sequence will be inserted into our organism, S.cerevisiae, with a plasmid that contains the yeast promoter,TEF.

Source

E2F1 is composed of a genomic sequence that encodes for E2F1 protein in humans. The amino acid sequence for E2F1 was determined from the website, http://www.uniprot.org/uniprot/Q01094 and converted into a DNA sequence using, https://www.idtdna.com/CodonOpt.

References

1)Describes the function of E2F1: http://atlasgeneticsoncology.org/Genes/E2F1ID40382ch20q11.html 2)Characterizes the relationship between E2F1 and p14ARF: http://www.ncbi.nlm.nih.gov/pubmed/12695664 and http://www.nature.com/onc/journal/v22/n32/full/1206659a.html