Regulatory

Part:BBa_K782072

Designed by: Fedja Pavlovec   Group: iGEM12_Slovenia   (2012-09-24)
Revision as of 22:32, 26 September 2012 by Fedjapavlovec (Talk | contribs)

12x[TALD] operator

  • TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).
  • DNA binding sites for individual TAL effectors are indicated with square brackets [ ].


Introduction

TAL effectors (TALEs) are able to specifically bind to the DNA by recognizing base pairs on a DNA molecule. With TAL regulators we are able to control expression of a desired gene. Efficient transcription control can be achieved by binding of multiple TAL regulators upstream of promoter.

Our construct contain twelve specific binding sites for TALD (Figure 1).

Single binding sequence for TALD is: TCGTCCAATAGCTTCTC


12xD.png

Figure 1. Shematic representation of the construct.


  • Binding sites for TAL effectors were ordered from IDT.


Characterization

Construct was later used for constructing different reporter plasmids to show specific binding of TALD:KRAB/TALD:VP16.

Results:


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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 214
    Illegal AgeI site found at 446
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None