Coding
frr

Part:BBa_K1668002:Design

Designed by: Xintian Xu   Group: iGEM15_ZJU-China   (2015-09-08)
Revision as of 09:44, 11 September 2015 by XuXintian (Talk | contribs) (→‎References)


frr (from Streptomyces avermitilis, increasing avermectin production)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 165
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 337


Design Notes

By PCR with primers frr1 and frr2 shown below, we added the standard prefix and suffix at both ends of the frr sequence. We used seamless assembly as our assembly method. Therefore, there are no scar between prefix sequence, frr, and suffix sequence.
frr1 (F, 5’-3’): GAATTCGCGGCCGCTTCTAGATGCGCGGGTACGTC
frr2 (R, 5’-3’): TGCAGCGGCCGCTACTAGTATTATTACATCAAGGTCGCC

Source

The frr gene was amplified by PCR with genomic DNA of S. avermitilis ATCC31267 strain as template.

References

Li, L., et al. (2010). "Overexpression of ribosome recycling factor causes increased production of avermectin in Streptomyces avermitilis strains." J Ind Microbiol Biotechnol 37(7): 673-679.