Composite

Part:BBa_K1365007:Design

Designed by: Sandra Mous   Group: iGEM14_Groningen   (2014-09-24)


NisF, NisE and NisG


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'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCAGGTAAAAATTCAAAA-3' reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTATCTAATCTTTTTTT-3' to PCR NisF, NisE and NisG from the genome and add the BioBrick prefix and suffix.

The illegal PstI site at position 186, illegal EcoRI site at postion 336 and illegal XbaI site at position 349 were removed using primers as well, fusing the resulting PCR fragments together again with a Gibson assembly.



Source

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

References