Part:BBa_K5142042
Reverse complementary GFP cds
This part contains the reverse complementary sequence of the coding sequence of green fluorescent protein (GFP). In our project, this part was inserted to the downstream of reverse poxviral H5 promoter (BBa_K5142012), and the expressed GFP can serve as an indicator of the successful transfection of the plasmid to cells infected by vaccinia virus and proximity of the plasmid to viral genome.
This part was created by PCR amplification according to the GFP coding region of the plasmid pCAGGS-GFP/A20 (GenBank ID: LT727038.1).
To prevent potential readthrough transcription when inserted into vaccinia virus genome, the GFP was transcribed in the direction reverse to the adjacent viral genes (e.g., A26L, A27L and A28L) which was driven by poxviral H5 promoter. Thus, a reverse complementary sequence of GFP cds should be added to the construct for virus engineering.
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 740 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal SpeI site found at 741
Illegal PstI site found at 755
Illegal NotI site found at 7
Illegal NotI site found at 748 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
Illegal BamHI site found at 215 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 741 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal XbaI site found at 16
Illegal SpeI site found at 741
Illegal PstI site found at 755 - 1000COMPATIBLE WITH RFC[1000]
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