Coding

Part:BBa_K5142038:Design

Designed by: Yuchen Liang   Group: iGEM24_HunanU   (2024-09-30)


Reverse complementary GFP cds


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 740
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal BamHI site found at 215
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 741
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

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, the a reverse complementary sequence of GFP cds should be added to the construct for virus engineering.


Source

This part was created by PCR amplification according to the GFP coding region of the plasmid pCAGGS-GFP/A20 (GenBank ID: LT727038.1).

References