Part:BBa_K3063041

Strand 4 (EpCam) for in vivo synthesis of DNA nanostructure

The biobrick design allows the in vivo production of Strand 4 ssDNA of DNA nano drug carrier with EpCam (cancer stem cell surface marker aptamer) attached at the 5' end.

The biobrick contains a promoter, a single strand DNA(ssDNA) production region, reverse transcriptase binding site (HTBS) and also a terminator. The promoter, ssDNA, HTBS site were constructed in a seamless way to allow correct ssDNA sequence after reverse transcription. The biobrick is used together with HIV reverse transcriptase (HIVRT) and Murine Leukemia Reverse Transcriptase (MLRT) co-expressed in E. Coli cells. It allows in vivo-synthesis of ssDNA due to the reverse transcription, and also RNA template degradation function of reverse transcriptases. The ssDNA produced could anneal to produce DNA nanostructure in vivo.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 13
  Illegal NheI site found at 36
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]