Part:BBa_K1336005:Design

Antisense for octaprenyl diphosphate synthase (ispB gene)

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

When designing PCR primers to change the reverse complement sequence of the ispB gene into the BioBrick format, we truncated the fragment to avoid an illegal PstI restriction site (and so no extra primers were required for site-directed mutagenesis to remove illegal sites). The specific primers were:

iGM PRE ispB RNAi: 5’ - GTTTCTTC GAATTC GCGGCCGC T TCTAGA G TTA ACG ATC GCG TTG AAC AGC G – 3’
iGM SUF REV ispB RNAi:5’–GT TTC TTC CTG CAG CGG CCG CTA CTA GTA ATG AAT TTA GAA AAA ATC AAT GAG–3’

Source

The PCR product was obtained from genomic E. coli sequence, via colony boil.