Composite

Part:BBa_K1365006:Design

Designed by: Sandra Mous   Group: iGEM14_Groningen   (2014-09-24)
Revision as of 02:37, 18 October 2014 by S.Mous (Talk | contribs) (→‎Design Notes)


NisR and NisK


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

Used primers:

forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATAAAATTTTAATAGT-3'

reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTTTTTTATTTTTA-3'

to PCR NisR and NisK from the genome and add the BioBrick prefix and suffix.


The GTG start codon of NisR was changed into a ATG, as this is standard with most BioBricks.


During the making of this BioBrick two mutations occured in the gene. At residue 199 of NisR a GAA was changed to a GAG, not changing the amino acid at this position. At residue 300 of NisK an ACA was changed to an ATA, changing a threonine to an isoleucine.

Source

Obtained by PCR on the genome of Lactococcus lactis NZ9700, a nisin producing strain.

References

Trmčić, A. et al. (2011) Complete nisin A gene cluster from Lactococcus lactis M78 (HM219853) – obtaining the nucleic acid sequence and comparing it to other published nisin sequences. Genes Genom. 33: 217-221