Coding

Part:BBa_K142040:Design

Designed by: Julius Rabl   Group: iGEM08_ETH_Zurich   (2008-10-16)
Revision as of 21:13, 16 October 2008 by Jrabl (Talk | contribs)

ribosome modulation factor (RMF)


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 55
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

RMF was amplified from E. coli, therefore sequence optimization was unnecessary. The RMF gene does not contain any restriction sites for EcoRI, XbaI, SpeI or PstI and could be turned into a BioBrick without further modification.


Source

The nucleotide sequence of RMF from Escherichia coli MG1655 was obtained from GenBank and used for primer design. Amplification from DH5-alpha genomic DNA template and addition of BioBrick standard prefix and suffix RMF was performed by PCR using the following primers:

forward primer 5'-cgcggaattcgcggccgcttctagatgaagagacaaaaacgagatcgcctgg

reverse primer 5'-cgcgctgcagcggccgctactagtattattaggccattactaccctgtccgc

The PCR product was subcloned into vector pSB1A7.

References