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

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

Revision as of 05:34, 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]


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

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


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.