Coding

Part:BBa_K1607010:Design

Designed by: Jiazheng Xing   Group: iGEM15_Shenzhen_SFLS   (2015-08-23)
Revision as of 05:31, 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]


The termination codon: 

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

Source

This coding sequence was synthesized by LCR assmebly using 36 oilgos

AATTCCGattgtgtcc R0 ggacacaatCGGAATTCATGGGTTCTACTCAAG F0 CATATCAGTACCAGTACAAACTTGAGTAGAACCCATG R16 TTTGTACTGGTACTGATATGAAGTTGAGATTGCCAG F33 AATGAGTTTCTGGAGAAGCTGGCAATCTCAACTT R53 CTTCTCCAGAAACTCATTTGGATATGTTGAGACATTT F69 CTTGACAACCTTGGTACAAATGTCTCAACATATCCA R87 GTACCAAGGTTGTCAAGTTGTTCAAGGTAACTTGG F106 TGGCAAGTAAGTCAATTCCAAGTTACCTTGAACAA R123 AATTGACTTACTTGCCAACTAACGCTTCTTTGTC F141 TCTTGAATATCTTGCAAAAAAGACAAAGAAGCGTTAGT R158 TTTTTTGCAAGATATTCAAGAAGTTCAAGGTTACGTTT F175 CTTGGTTATGAGCAATCAAAACGTAACCTTGAACT R196 TGATTGCTCATAACCAAGTTAGACAAGTTCCATTGC F213 CTCTAACAATTCTCAATCTTTGCAATGGAACTTGTCTAA R231 AAAGATTGAGAATTGTTAGAGGTACTCAATTGTTTGAAGAT F249 CAGCCAAAGCGTAGTTATCTTCAAACAATTGAGTAC R270 AACTACGCTTTGGCTGTTTTGGATAACGGTGATC F290 GGAGTAGTGTTGTTCAATGGATCACCGTTATCCAAAA R306 CATTGAACAACACTACTCCAGTTACTGGTGCTTCT F324 CTCTCAAACCACCTGGAGAAGCACCAGTAACT R343 CCAGGTGGTTTGAGAGAATTGCAATTGAGATCTTT F359 CCCTTCAAAATTTCAGTCAAAGATCTCAATTGCAATT R375 GACTGAAATTTTGAAGGGTGGTGTTTTGATTCAAAG F394 ACACAATTGTGGGTTTCTTTGAATCAAAACACCA R412 AAACCCACAATTGTGTTACCAAGATACTATTTTGTGG F430 TGTTCTTATGAAAAATATCCTTCCACAAAATAGTATCTTGGTA R446 AAGGATATTTTTCATAAGAACAACCAATTGGCTTTGAC F467 GATCTGTTAGTATCAATCAAAGTCAAAGCCAATTGGT R489 TTTGATTGATACTAACAGATCCAGAGCTTGTCATCCA F505 CTTACACATTGGAGAACATGGATGACAAGCTCTG R526 TGTTCTCCAATGTGTAAGGGTTCCAGATGTTGGG F542 CAATCTTCAGAAGATTCACCCCAACATCTGGAACC R560 GTGAATCTTCTGAAGATTGTCAATCTTCTAGACCca F576 ggacacgtaaatcactattgGGTCTAGAAGATTGA R595 atagtgatttacgtgtcc R612

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

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.