Difference between revisions of "Part:BBa K1607011:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1607011 short</partinfo>
 
<partinfo>BBa_K1607011 short</partinfo>
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===Design Notes===
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===Source===
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design information will be provided soon
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The scFv CDS was synthesized by LCR assmebly using 36 oilgos
  
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[[see oligos and protocols]]
  
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The eGFP CDS was obtained using PCR.
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Primers:
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ggaggcggaggcagcggtggcggtggaagcATGGTGAGCAAGGGCga (eGFP-F)
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GGTCTAGAGGCTTGTACAGCTCGTCCAT (eGFP-R)
  
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===Source===
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The eGFP CDS and the scFv CDS were connected by SOE PCR
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Primers:
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LCR assembly and PCR.(detailed description will be provided soon)
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step1:
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CGGAATTCatgGGTTCTACTCAAG (SCFV-F)
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gctgcctccgcctccagaaccgccaccgccAGATTGACAATCTTCagaaga (SCFV-R)
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ggaggcggaggcagcggtggcggtggaagcATGGTGAGCAAGGGCga (eGFP-F)
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GGTCTAGAGGCTTGTACAGCTCGTCCAT (eGFP-R)
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step2:
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SCFV-F and eGFP-R
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Standard Biobrick prefix and suffix were then added to the sequence by PCR, replacing the EcoRI site and XbaI site which were originally designed for yeast expression.
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Primers:
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GAATTCGCGGCCGCTTCTAGatgGGTTCTACTCAAGttt (F)
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TACTAGTAGCGGCCGCTGCAGCTTGTACAGCTCGTCCAT(R)
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===Design Notes===
  
 
===References===
 
===References===

Revision as of 13:16, 17 September 2015

The SCFV of anti-p185her2/neu antibody chA21 linked with EGFP by a (Gly4Ser)3 linker


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

The scFv CDS was synthesized by LCR assmebly using 36 oilgos

see oligos and protocols

The eGFP CDS was obtained using PCR. Primers: ggaggcggaggcagcggtggcggtggaagcATGGTGAGCAAGGGCga (eGFP-F) GGTCTAGAGGCTTGTACAGCTCGTCCAT (eGFP-R)

The eGFP CDS and the scFv CDS were connected by SOE PCR Primers: step1: CGGAATTCatgGGTTCTACTCAAG (SCFV-F) gctgcctccgcctccagaaccgccaccgccAGATTGACAATCTTCagaaga (SCFV-R) ggaggcggaggcagcggtggcggtggaagcATGGTGAGCAAGGGCga (eGFP-F) GGTCTAGAGGCTTGTACAGCTCGTCCAT (eGFP-R) step2: SCFV-F and eGFP-R

Standard Biobrick prefix and suffix were then added to the sequence by PCR, replacing the EcoRI site and XbaI site which were originally designed for yeast expression. Primers: GAATTCGCGGCCGCTTCTAGatgGGTTCTACTCAAGttt (F) TACTAGTAGCGGCCGCTGCAGCTTGTACAGCTCGTCCAT(R)

Design Notes

References