Part:BBa_K782081

12x[TALD] operator_CMV promoter_2xBFP:NLS

TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).

Introduction

Our construct contain TALD binding sites that 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.

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 465
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 214
  Illegal AgeI site found at 446
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1141
  Illegal BsaI.rc site found at 1797
  Illegal BsaI.rc site found at 2502