Composite

Part:BBa_K2282009:Design

Designed by: Eliott LAFON   Group: iGEM17_IONIS-PARIS   (2017-10-02)


BBa_K592009 (AmilCP) consitutive expression system with BBa_K2282001 (CspA promoter and Up element).


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Finding the UP element and the promoter was tricky and we needed certain amount of time to make sure we did not miss the sequence in the NCBI GenBank.



Source

Go to: https://www.ncbi.nlm.nih.gov/gene/948070 and in the 5'UTR. The Up element is an "AT-rich sequence (-47 to -38) upstream of the CspA -35 region. If you look now on the Cold-shock protein A gene on this region, you will find the sequence. If you take the CspA gene, you can then go to nucleotide number 3719890 (ACGGTT) to have the beginning of the 5'UTR. Then you need to go further to 3719843 (AAAATAATTG). You will notice that the sequence is indeed located from -47 to -38 from the 5'UTR beginning and is AT rich. Sequences of the cold-induced elements are very close to the parts of the iGEM10_Mexico-UNAM-CINVESTAV that can be found here : http://partsregistry.org/Part:BBa_K328003


References