Difference between revisions of "Part:BBa K176157:Design"

 
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<partinfo>BBa_K176157 short</partinfo>
 
<partinfo>BBa_K176157 short</partinfo>
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Add LVA degradation tag (widely used in C0040, C0051, C0061...) to ccdB(K145151) C-terminal:
 
Add LVA degradation tag (widely used in C0040, C0051, C0061...) to ccdB(K145151) C-terminal:
  
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[K145151 without stop codon] GCT GCA AAC GAC GAA AAC TAC GCT TTA GTA GCT [TAA TAA]
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[K145151 without stop codon] GCT GCA AAC GAC GAA AAC TAC GCT TTA GTA GCT [TAA TAA]
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                              A  A  N  D  E  N  Y  A  L  V  A
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                            A  A  N  D  E  N  Y  A  L  V  A
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===Source===
 
===Source===

Latest revision as of 13:36, 14 September 2009

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