Composite

Part:BBa_K782082

Designed by: Fedja Pavlovec   Group: iGEM12_Slovenia   (2012-09-24)
Revision as of 22:12, 25 September 2012 by UrosZupancic (Talk | contribs)

12x[NicTAL] operator_CMV promoter_2xBFP:NLS


Introduction

Our construct contain twelve binding sites for NicTAL12 (binding site's sequence) that are cloned upstream of CMV promoter. Downstream of CMV is a double blue fluorescent protein with NLS domain, causing protein to migrate into the nucleus. BFP is a monomeric fluorescent protein with excitation maximum at 402 nm and emision maximum at 457 nm.

Nicbfp2.png

Figure 1: Schematic representation of the construct.


Characterization

Results: Specific TAL binding sites were further characterized with other reporter constructs.

BFP was obtained from Evrogen.

References

Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 464
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 16
    Illegal AgeI site found at 248
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1139
    Illegal BsaI.rc site found at 1795
    Illegal BsaI.rc site found at 2500


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Categories
Parameters
None