Coding

Part:BBa_K1607010:Design

Designed by: Jiazheng Xing   Group: iGEM15_Shenzhen_SFLS   (2015-08-23)
Revision as of 06:02, 24 August 2015 by VictorX (Talk | contribs) (→‎Source)

The coding sequence of the SCFV of anti-p185her2/neu antibody chA21


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 370
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Source

This coding sequence was synthesized by LCR assmebly using 36 oilgos

oligos and protocol

Design note

Isolation:

After the LCR assembly, we isolated the CDS using these two primers(with BioBrick Prefix in the fwd primer and Suffix in the rev primer):

fwd:GAATTCGCGGCCGCTTCTAGatgGGTTCTACTCAAGttt

rev:TACTAGTAGCGGCCGCTGCAGAGATTGACAATCTTCAGAAGAT

The termination codon:

Since it was originally designed for 3'his-tag vectors, this coding sequence does not have a termination codon.

References

[1]Zhou H, Zha Z, Liu Y, et al. Structural Insights into the Down-regulation of Overexpressed p185her2/neu Protein of Transformed Cells by the Antibody chA21[J]. Journal of Biological Chemistry, 2011, 286(36): 31676-31683.