Part:BBa_K5142037

Reverse poxviral H5 promoter

This part includes a DNA fragment derived from the promoter of H5 gene of vaccinia virus. This part can drive the transcription of its downstream element with high efficiency ONLY in poxviral factory, a specific region in cytoplasm for poxvirus replication stablished during poxvirus infection in cells, which means that this promoter cannot be used to drive transcription in nuclei or mitochondria. It must be noted that this part is a reverse promoter and the downstream coding sequence should be added to its 5’ end with in a reverse complementary form. In this project, we used this promoter to drive the expression of GFP on the plasmid for vaccinia virus recombination, which could indicate the successful transfection of the plasmid to cells infected by vaccinia virus and proximity of the plasmid to viral genome.

Usage and Biology

This part was synthesized according to the sequence of the promoter of H5 gene in vaccinia virus genome (GenBank ID: NC_006998.1, 91, 764 to 91,832 bp).

The 3’-end nucleotide of the promoter and the 5’-end nucleotide of Biobrick suffix are both T, thus we delete one T at the 3’ end of the promoter to reduce the size of the part.

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 1
  Illegal suffix found in sequence at 130
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1
  Illegal SpeI site found at 131
  Illegal PstI site found at 145
  Illegal NotI site found at 7
  Illegal NotI site found at 138
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1
  Illegal XhoI site found at 48
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 1
  Illegal suffix found in sequence at 131
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 1
  Illegal XbaI site found at 16
  Illegal SpeI site found at 131
  Illegal PstI site found at 145
  Illegal NgoMIV site found at 33
  
• 1000
  COMPATIBLE WITH RFC[1000]