DNA

Part:BBa_K1951000:Design

Designed by: Claire raynaud, Yoann Santin, Louis Givelet   Group: iGEM16_Aix-Marseille   (2016-10-10)

Sequence optimization

Snapgene has been used to remove the forbidden sites by changing in the substitution which didn't change the amino acid. The sequence has been sythetised by IDT[1].

Codon optimization for Escherichia coli.

We obtimised codon for Escherichia coli. Codon optimization is a technical used to improve the protein expression in living organism by increasing the translational efficiency of gene of interest [1-4, 6-13, 15-18, 20-28]. This biobrick codon optimised increase the functionality of gene. To process it, we use codon optimization IDT software[2]. If you use this biobrick in Escherichia coli, you can be sure that the protein produced will be highly expressed and well solubilised.

Prefix and suffix addition

Prefix and suffix subsequences (containing restriction site EcoRI, XbaI and SpeI PstI respectively) have been added by a SLIC method with the following oligos :

desA slic forward cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGATGCGCTCGCACTTGCTT
desA slic reverse ttgcccttttttgccggaCTGCAGCGGCCGCTACTAGTATTATTAGGAGGCACGGTC
.

SequenceAndFeatures


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 346
    Illegal NgoMIV site found at 1393
  • 1000
    COMPATIBLE WITH RFC[1000]

Source

This part comes from the genome sequencing of Streptomyces coelicolor. The following website enabled to find the gene reference and its chromosomal position. http://strepdb.streptomyces.org.uk/cgi-bin/dc3.pl?accession=AL645882&serial=2769&width=900&start=3031362&end=3041362&iorm=map

The second website below is the source of the DNA sequence we have used. http://www.ncbi.nlm.nih.gov/nuccore/NC_003888.3

References

  1. ↑ http://eu.idtdna.com/site
  2. ↑ https://eu.idtdna.com/CodonOpt