Difference between revisions of "Part:BBa K1607010:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1607010 short</partinfo> | <partinfo>BBa_K1607010 short</partinfo> | ||
<partinfo>BBa_K1607010 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1607010 SequenceAndFeatures</partinfo> | ||
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===Source=== | ===Source=== | ||
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atagtgatttacgtgtcc | 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): | 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): | ||
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rev:TACTAGTAGCGGCCGCTGCAGAGATTGACAATCTTCAGAAGAT | rev:TACTAGTAGCGGCCGCTGCAGAGATTGACAATCTTCAGAAGAT | ||
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| + | '''The termination codon:''' | ||
| + | Since it was originally designed for 3'his-tag vectors, this coding sequence does not have a termination codon. | ||
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===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 05:39, 24 August 2015
The coding sequence of the SCFV of anti-p185her2/neu antibody chA21
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 370
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.