Composite

Part:BBa_K782038

Designed by: Fedja Pavlovec   Group: iGEM12_Slovenia   (2012-09-20)

10x[TALA] operator_CMV promoter_TALB:NLS:KRAB


  • TALB and TALA labels represents TAL effectors 1297 and 1257 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 10 TALA 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.

SVN12 10A TBK.png

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 914
    Illegal BamHI site found at 1546
    Illegal BamHI site found at 4016
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 341
    Illegal NgoMIV site found at 701
    Illegal AgeI site found at 206
    Illegal AgeI site found at 541
    Illegal AgeI site found at 566
    Illegal AgeI site found at 901
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 4382
    Illegal SapI.rc site found at 4346


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