Composite
Part:BBa_K1365007:Design
Designed by: Sandra Mous Group: iGEM14_Groningen (2014-09-24)
NisF, NisE and NisG
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.