Part:BBa_K1172904:Design

Rnase Ba (Barnase) from Bacillus amyloliquefaciens

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

Amplification with specfic Gibson-Primer

• Amplification of the RNase Ba (Barnase):
  

Barnase_fwd (50 bp)

5'-ATTCGCGGCCGCTTCTAGAGATGGCAGCACAGGTTATCAACACGTTTGAC-3'

Barnase_rev (44)

5'-TGCAGCGGCCGCTACTAGTATTATCTGATTTTTGTAAAGGTCTG-3'

• Amplification of the pSB1C3-Backbone:
  

Barnase_suf (47)

5'-CAGACCTTTACAAAAATCAGATAATACTAGTAGCGGCCGCTGCAGTC-3'

Barnase_pre (53)

5'-GTCAAACGTGTTGATAACCTGTGCTGCCATCTCTAGAAGCGGCCGCGAATTCC-3'

• Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].

Source

• Genomic DNA-Isolation from Bacillus amyloliquefaciens DMSZ7.
  • Isolation of the whole B. amyloliquefaciens genome, see [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Whole_Genome_Isolation here]