Part:BBa_K4855003

J23101 - T7 - medium RBS - blaNDM2&5 CDS

This composite part comprises of J23101 - T7 - medium RBS - blaNDM-2&5. It was conceptualized by the iGEM Bulgaria 2023 team as a construct intended for in vivo T7-driven mutagenesis for directed evolution purposes. It was cloned in the low copy number SC101 ori-based vector pSB4K5 and used with the dual gene-specific mutator system capable of introducing transition mutations at consistent frequencies within a target sequence in vivo that was available for ordering from Addgene. (Seo D, Koh B, Eom GE, Kim HW, Kim S. A dual gene-specific mutator system installs all transition mutations at similar frequencies in vivo. Nucleic Acids Res. 2023 Jun 9;51(10):e59. doi: 10.1093/nar/gkad266. PMID: 37070179; PMCID: PMC10250238.).

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]