Part:BBa_K1681000

• Part Name: PL lac O-1 SCRIBE(kanR)ON
• Short Description: IPTG-inducible kanamycin resistance SCRIBE cassette
• Long Description: SCRIBE(kanR)ON is a cassette coding for the RNA and proteins needed to restore kanamycin resistance by synthesizing multicopy single-stranded DNA (msDNA) for site-specific recombination. The part consists of the IPTG-inducible promoter PL lac O-1, a retron coding for an untranslated RNA molecule and the open reading frame for reverse transcriptase, a RBS, the open reading frame for beta recombinase, and a terminator. The reverse transcriptase uses the msDNA (msd) region of the RNA molecule as a template and the msd RNA (msr) region of the RNA molecule as a primer to synthesize the msDNA. The beta recombinase inserts the kanamycin resistance segment of the msDNA into the host’s genome during DNA replication as an Okazaki fragment.
• Design Considerations: This part was designed to be used as a positive control for testing SCRIBE cassettes with different promoters and/ or custom DNA segments for site-specific recombination.

This graph shows our construct being induced. As you can see, the more IPTG added, the more recombination as a result. The graph is on a log log scale and follows an upward trend.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1267
  Illegal XhoI site found at 509
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]