Coding

Part:BBa_K1607010:Design

Designed by: Jiazheng Xing   Group: iGEM15_Shenzhen_SFLS   (2015-08-23)
Revision as of 05:41, 24 August 2015 by VictorX (Talk | contribs) (→‎Design note)

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

AATTCCGattgtgtcc

ggacacaatCGGAATTCATGGGTTCTACTCAAG

CATATCAGTACCAGTACAAACTTGAGTAGAACCCATG

TTTGTACTGGTACTGATATGAAGTTGAGATTGCCAG

AATGAGTTTCTGGAGAAGCTGGCAATCTCAACTT

CTTCTCCAGAAACTCATTTGGATATGTTGAGACATTT

CTTGACAACCTTGGTACAAATGTCTCAACATATCCA

GTACCAAGGTTGTCAAGTTGTTCAAGGTAACTTGG

TGGCAAGTAAGTCAATTCCAAGTTACCTTGAACAA

AATTGACTTACTTGCCAACTAACGCTTCTTTGTC

TCTTGAATATCTTGCAAAAAAGACAAAGAAGCGTTAGT

TTTTTTGCAAGATATTCAAGAAGTTCAAGGTTACGTTT

CTTGGTTATGAGCAATCAAAACGTAACCTTGAACT

TGATTGCTCATAACCAAGTTAGACAAGTTCCATTGC

CTCTAACAATTCTCAATCTTTGCAATGGAACTTGTCTAA

AAAGATTGAGAATTGTTAGAGGTACTCAATTGTTTGAAGAT

CAGCCAAAGCGTAGTTATCTTCAAACAATTGAGTAC

AACTACGCTTTGGCTGTTTTGGATAACGGTGATC

GGAGTAGTGTTGTTCAATGGATCACCGTTATCCAAAA

CATTGAACAACACTACTCCAGTTACTGGTGCTTCT

CTCTCAAACCACCTGGAGAAGCACCAGTAACT

CCAGGTGGTTTGAGAGAATTGCAATTGAGATCTTT

CCCTTCAAAATTTCAGTCAAAGATCTCAATTGCAATT

GACTGAAATTTTGAAGGGTGGTGTTTTGATTCAAAG

ACACAATTGTGGGTTTCTTTGAATCAAAACACCA

AAACCCACAATTGTGTTACCAAGATACTATTTTGTGG

TGTTCTTATGAAAAATATCCTTCCACAAAATAGTATCTTGGTA

AAGGATATTTTTCATAAGAACAACCAATTGGCTTTGAC

GATCTGTTAGTATCAATCAAAGTCAAAGCCAATTGGT

TTTGATTGATACTAACAGATCCAGAGCTTGTCATCCA

CTTACACATTGGAGAACATGGATGACAAGCTCTG

TGTTCTCCAATGTGTAAGGGTTCCAGATGTTGGG

CAATCTTCAGAAGATTCACCCCAACATCTGGAACC

GTGAATCTTCTGAAGATTGTCAATCTTCTAGACCca

ggacacgtaaatcactattgGGTCTAGAAGATTGA

atagtgatttacgtgtcc

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.