Part:BBa_K782064
12x[TALD] operator_CMV promoter_TALB:NLS:KRAB
- TALB and TALD labels represents TAL effectors 1297 and 1295 respectively from zebrafish experiments (Sander et al., 2011).
- DNA binding sites for individual TAL effectors are indicated with square brackets [ ].
Introduction
TAL effectors (TALEs) are bacterial plant pathogen transcription factors, that bind to DNA by specifically recognizing one base pair with a single tandem repeat in their DNA-binding domain. A tandem TALE repeat contains 33 to 35 amino acids, where the 12th and 13th amino acid, called a “repeat variable diresidue” (RVD), are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011).
This part contains 12 TALD binding sites, upstream of a CMV promoter (constitutive promoter for expression in mammalian cells). Downstream of the promoter there is a TAL repressor TALB:NLS:KRAB. This way we have a TAL effector under the control of another TAL effector, which allows us to create more complicated genetic circuits with more than one level of regulation.
Figure 1: Schematic representation of the construct.
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.
Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 659
Illegal BamHI site found at 1291
Illegal BamHI site found at 3761 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 408
Illegal AgeI site found at 640 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 4127
Illegal SapI.rc site found at 4091
| None |