Translational_Unit

Part:BBa_K415117:Design

Designed by: Grant Robinson   Group: iGEM10_MIT   (2010-07-11)

RBS Weak : pVIII-Fos


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 410
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 500
    Illegal BsaI.rc site found at 1107


Design Notes

The sequence for Fos is extensive at 1143 bp; therefore, designing the sequence as a simple primer is prohibitively costly. We instead constructed the sequence using PCR SOEing; in between the parts is a [pentaglycine linker bridge (GGAGGAGGAGGAGGA) or glycine-serine bridge (GGAGGAGGAGGAAGU).]

In addition, the Fos gene underwent site-directed mutagenesis at nucleotides 51 (C to T), 222 (G to A), and 453 (A to C) in order to remove three PstI sites.

Source

The source of the weak RBS is part BBa_B0031. The source of the pVIII protein is the genomic sequence of the M13KE bacteriophage, and the Fos protein can be found in the PubMed nucleotide archives, specifically at the following link.

http://www.ncbi.nlm.nih.gov/nuccore/31560587?from=140&to=1282&report=gbwithparts

References