Coding

Part:BBa_K1365005:Design

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


NisP


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'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAAAAAAATACTAGGTTT-3' reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATCAATTTTTAGTCTTTC-3' to PCR NisP from the genome and add the BioBrick prefix and suffix.

Changed the start codon from a GTG to an ATG.

The illegal PstI sites at position 225 and the illegal EcoRI sites at postion 347 and 1657 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