Coding

Part:BBa_K1111014:Design

Designed by: Sandra Elisabeth Chaudron, Caroline Desmurget and Mareike Apelt   Group: iGEM13_EPF_Lausanne   (2013-09-19)
Revision as of 21:32, 2 October 2013 by Sandelisa90 (Talk | contribs) (→‎Design Notes)

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]

Gibson Assembly Design

Insert: we amplified the streptavidin coding from the Biobrick BBa_K283010 designed by iGEM 2009 group HKU-HKBU.
Backbone: Starting from the biobrick BBa_K523013 (INP fused with YFP), we amplified the whole sequence except the EYFP and added gibson overhang complementary to streptavidin ends.

Primers

Streptavidin iGEM PCR :
5' ATGGCTGAAGCTGGTATCACC 3'
5' TTAGGAAGCAGCGGACGGTTTAAC 3'

BBa_K523013 PCR :
Fw: 5' ACCAAAGTTAAACCGTCCGCTGCTTCTAACATATCATAACGGAGTGATCGCAATG 3'
Rev: 5' CCAGGTGCCGGTGATACCAGCTTCAGCCATAGATCCCGCCACGCTGCT 3'

References and Acknowledgements

Thanks to the iGEM group that designed this Biobrick.

Source

Can be expressed in Escherichia Coli.

References and acknowledgements

Thanks to the Mark Howarth laboratory at Oxford university Dept. of Biochemistry (link) for providing us with the streptavidin cloning plasmid. Thanks also to the iGEM team of Edinburgh 2011 that designed the Biobrick BBa_K523013.