Part:BBa_K4816004

nirS primer reverse

• Primer design

This primer was designed with a region to flank the 3' terminus of the gene NirS in Paraccocus denitrificans PD1222 part of the chromosome 1 (NCBI ID:CP000489.1) located between base pairs (2503999 and 2505789). The primer contains the addition of the restriction site BlpI and 6 bases to allow the cut of the restriction endonuclease. The introduction of the unique restriction site BlpI into the gene NirS allows the precise introduction of NirS in the plasmid vector pet 23 a (+) for protein expression and further purification. The primer contains 16 bp DNA binding region. Due to the presence of XhoI inside the gene of interest, another unique restriction site was chosen BlpI, due to this change his tag was added to the C terminal for Protein purification.

 GTCACTGCTNAGCCTAGTGGTGATGGTGATGATGATAGGTGTCGGTCATA
       *****+++********************----------------
       BlpI S.C       His-Tag      DNA binding region
 
 S.C= Stop Codon

Sequence and Features

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]