Part:BBa_K782069
10x[TALA+TALB] operator
- TALA and TALB labels represents TAL effector 1257 and 1297 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 ten binding sites for TALA and another ten for TALB. Single binding site is combined from 18 base pairs recognised by specific TAL. TALA and TALB binding sites are adjacent to each other making impossible for both of them to bind at once.
Single binding site sequence for TALA: TTTACTGCTGCTCCCGCT
Single binding site sequence for TALB: TCTTCCGTTTCCACATCT
Figure 1: Schematic representation of the construct.
- Binding sites for TAL effectors were ordered from GeneArt.
Characterization
Construct was later used for constructing different reporter plasmids to show specific binding and competition between TALA and TALB.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]
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