Reporter

Part:BBa_K782078

Designed by: Fedja Pavlovec   Group: iGEM12_Slovenia   (2012-09-24)
Revision as of 22:11, 25 September 2012 by UrosZupancic (Talk | contribs)

10x[TALA+TALB] operator_CMV promoter_2xBFP:NLS

TALA and TALB labels represents TAL effector 1257 and 1297 from zebrafish experiments (Sander et al., 2011).

Introduction

Our construct contain TALA and TALB binding sites, which are adjacent to each other. Because of steric interference two different TAL regulators are unable to bind to its binding sites at once. TAL binding sites 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.

ABbfp2.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 730
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 149
    Illegal NgoMIV site found at 514
    Illegal AgeI site found at 12
    Illegal AgeI site found at 352
    Illegal AgeI site found at 377
    Illegal AgeI site found at 717
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1406
    Illegal BsaI.rc site found at 2062
    Illegal BsaI.rc site found at 2767


[edit]
Categories
Parameters
None