Difference between revisions of "Part:BBa K1111014:Design"
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<partinfo>BBa_K1111014 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1111014 SequenceAndFeatures</partinfo>
  
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==Design Notes==
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To assemble this part, we asked for the Biobrick BBa_K283010 from iGEM 2009 group HKU-HKBU ([https://parts.igem.org/Part:BBa_K283010 Biobrick page]).
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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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Gibson Assembly
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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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Overlapping PCR of BBa_K523013 :
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5' accaaagttaaaccgtccgctgcttcctaacatatcataacggagtgatcgcaatg 3'
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5' ccaggtgccggtgataccagcttcagcCATagatcccgccacgctgct 3'
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==References and Acknowledgements==
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Thanks to the iGEM group that designed this Biobrick.
  
  

Revision as of 11:12, 2 October 2013

Ice Nucleation Protein fused to Streptavidin BBa_K283010


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1649
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    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 asked for the Biobrick BBa_K283010 from iGEM 2009 group HKU-HKBU (Biobrick page). 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 iGEM group that designed this Biobrick.


Source

Can be expressed in Escherichia Coli

References