Difference between revisions of "Part:BBa K1111012:Design"
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==References and Acknowledgements== | ==References and Acknowledgements== | ||
| − | Thanks to the Mark Howarth lab, Biochemistry Dpt in Oxford | + | Thanks to the Mark Howarth lab, Biochemistry Dpt in Oxford for providing us with the streptavidin cloning plasmid. |
Revision as of 17:51, 2 October 2013
Ice Nucleation Protein fused to Streptavidin Alive
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1727
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1649
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1727
- 25INCOMPATIBLE 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 - 1000COMPATIBLE 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 with the streptavidin cloning plasmid.