Difference between revisions of "Part:BBa K1607010:Design"

(Design note)
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This coding sequence was synthesized by LCR assmebly using 36 oilgos
 
This coding sequence was synthesized by LCR assmebly using 36 oilgos
  
AATTCCGattgtgtcc                    
+
[[see oligos and protocols]]
 
+
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===
 
===Design note===

Latest revision as of 06:02, 24 August 2015

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

see oligos and protocols

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.