Difference between revisions of "Part:BBa K1111014:Design"

Revision as of 21:30, 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

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.

@@ Line 9: / Line 9: @@
 ==Primers==
-Streptavidin Alive PCR :
+Streptavidin iGEM PCR :
 <br>5' <font color = #0000FF>ATGGCTGAAGCTGGTATCACC</font> 3'
 <br>5' <font color = #0000FF>TTAGGAAGCAGCGGACGGTTTAAC</font> 3'