Coding
ccdB-LVA

Part:BBa_K176157:Design

Designed by: Danqian Liu, Zongxiao He, Hao Jiang   Group: iGEM09_USTC   (2009-09-14)
Revision as of 13:36, 14 September 2009 by Horigen (Talk | contribs) (→‎Design Notes)

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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.

References