Reporter

Part:BBa_K782081

Designed by: Fedja Pavlovec   Group: iGEM12_Slovenia   (2012-09-24)
Revision as of 11:02, 26 September 2012 by LucijaK (Talk | contribs) (Characterization)

12x[TALD] operator_CMV promoter_2xBFP:NLS


TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).

Introduction

Our construct contain TALD binding sites 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.

Dbfp2.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 465
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 214
    Illegal AgeI site found at 446
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1141
    Illegal BsaI.rc site found at 1797
    Illegal BsaI.rc site found at 2502


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