Coding

Part:BBa_K1949020:Design

Designed by: Kazuki Fujisawa   Group: iGEM16_Tokyo_Tech   (2016-10-06)
Revision as of 07:59, 11 October 2016 by Yoshio (Talk | contribs)


yafN


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 AgeI site found at 78
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

yafN(BBa_K1949020) was gained by PCR using E. coli MG1655 genomic DNA and the following primers.

fwd:5’-aaatctagatgcatcgaattctcgctgaaaaatcg-3’

rev:5’-cccctgcagcggccgctactagtattattattccttaaagtcattgaaatcatcatccgtc-3’

yafN gene fragment amplified by PCR contains XbaI site at N-terminal and SpeI and PstI site at C-terminal. Both terminals were cut by respective restriction enzymes and the fragment used for ligation with iGEM plasmid vectors.


Material and method

Source

From E.coli MG1655.

Material and method

References

[1] Zhang Y, Yamaguchi Y, Inoue M. Characterization of YafO, an Escherichia coli Toxin. J Biol Chem. 2009 Sep; 284(38): 25522–25531.

[2] Dalsgaard M, Jørgensen M, Gerdes K. Three new RelE-homologous mRNA interferases of Escherichia coli differentially induced by environmental stresses. Mol Microbiol. 2010 Jan; 75(2): 333–348.

[3] Singletary LA, Gibson JL, Tanner EJ, McKenzie GJ, Lee PL, Gonzalez C, et all. An SOS-Regulated Type 2 Toxin-Antitoxin System. J Bacteriol. 2009 Dec;191(24):7456-65.