Part:BBa_K176157:Design

ccdB-LVA coding sequence

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 216
  

Design Notes

Add LVA degradation tag (widely used in C0040, C0051, C0061...) to ccdB(K145151) C-terminal:

[K145151 without stop codon] GCT GCA AAC GAC GAA AAC TAC GCT TTA GTA GCT [TAA TAA]
                              A   A   N   D   E   N   Y   A   L   V   A

Source

PCR from K145151.

Forward Primer: VF2(G00100)

Reverse Primer: GTTT ACTAGT A TTATTA AGC TAC TAA AGC GTA GTT TTC GTC GTT TGC AGC tattccccagaacatcaggtta

Digest with EcoRI and SpeI, ligate into pSB1A3.