Part:BBa_K3878020

T7 promoter +hsa-miRNA-21_5p + T7 terminator

This part enables the expression of human microRNA-21-5p (hsa-miR-21-5p) via T7 RNA polymerase activity. It is composed of the BglII restriction site at its 5’ end and BamHI restriction site at its 3’ end. The hsa-miR-21-5p sequence is preceded by the T7 promoter and is followed by the T7 terminator.

Usage and Biology

This part is flanked by BglII and BamHI restriction sites for assembly in a receiving vector via molecular cloning using BglII and BamHI restriction enzymes as described below (Fig. 1).

Figure 1. Principle of the insertion of hsa-miR-141-5p fragment in a plasmid. Created with BioRender.

This part, when expressed in a cell or cell-free system containing T7 RNA polymerase, will produce a hsa-miRNA-21(5p. A random nucleotide sequence has been designed between the T7 terminator and the BamHI restriction sites to lengthen the size of the part. It enables easy visualization on an agarose gel. E. coli BL21 DE3 strain can be used to express this part in vivo.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 194
  Illegal SpeI site found at 230
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 230
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1
  Illegal BamHI site found at 247
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 194
  Illegal SpeI site found at 230
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 194
  Illegal SpeI site found at 230
  
• 1000
  COMPATIBLE WITH RFC[1000]