Difference between revisions of "Part:BBa K1111012:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1111012 short</partinfo>
 
<partinfo>BBa_K1111012 short</partinfo>
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<partinfo>BBa_K1111012 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1111012 SequenceAndFeatures</partinfo>
  
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==Design Notes==
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To assemble this part, we orderd a plasmid containing the streptavidin alive.
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We amplified only the coding sequence of streptavidin of the plasmid by PCR. Also, we did a PCR with overlapping ends corresponding to streptavidin on the '''Biobrick BBa_K523013''' (INP fused with YFP). This PCR was keeping all the plasmid including RBS and Lac promoter but excluding YFP. The linearized plasmid and the insert were then assembled by Gibson assembly.
  
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===Design Notes===
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==Primers==
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Remove YFP sequence form BBa_K523013
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PCR of streptavidin alive :
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5' ATGGCTGAAGCTGGTATCACC 3'
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5' TTAGGAAGCAGCGGACGGTTTAAC 3'
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===Source===
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Gibson Assembly of parts
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Overlapping PCR of BBa_K523013 :
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5' accaaagttaaaccgtccgctgcttcctaacatatcataacggagtgatcgcaatg 3'
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5' ccaggtgccggtgataccagcttcagcCATagatcccgccacgctgct 3'
  
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===References===
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==References and Acknowledgements==
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Thanks to the Mark Howarth lab, Biochemistry Dpt in Oxford, for providing us the needed plasmid.

Revision as of 10:58, 2 October 2013

Ice Nucleation Protein fused to Streptavidin Alive


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1727
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1649
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1727
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1727
    Illegal NgoMIV site found at 1036
    Illegal AgeI site found at 1691
    Illegal AgeI site found at 1742
  • 1000
    COMPATIBLE WITH RFC[1000]

Design Notes

To assemble this part, we orderd a plasmid containing the streptavidin alive. We amplified only the coding sequence of streptavidin of the plasmid by PCR. Also, we did a PCR with overlapping ends corresponding to streptavidin on the Biobrick BBa_K523013 (INP fused with YFP). This PCR was keeping all the plasmid including RBS and Lac promoter but excluding YFP. The linearized plasmid and the insert were then assembled by Gibson assembly.

Primers

PCR of streptavidin alive : 5' ATGGCTGAAGCTGGTATCACC 3' 5' TTAGGAAGCAGCGGACGGTTTAAC 3'

Overlapping PCR of BBa_K523013 : 5' accaaagttaaaccgtccgctgcttcctaacatatcataacggagtgatcgcaatg 3' 5' ccaggtgccggtgataccagcttcagcCATagatcccgccacgctgct 3'

References and Acknowledgements

Thanks to the Mark Howarth lab, Biochemistry Dpt in Oxford, for providing us the needed plasmid.