Part:BBa_K782079

10x[TALA] operator_CMV promoter_2xBFP:NLS

TALA labels represents TAL effector 1257 from zebrafish experiments (Sander et al., 2011).

Introduction

Our construct contain TALA 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 720
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 147
  Illegal NgoMIV site found at 507
  Illegal AgeI site found at 12
  Illegal AgeI site found at 347
  Illegal AgeI site found at 372
  Illegal AgeI site found at 707
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1396
  Illegal BsaI.rc site found at 2052
  Illegal BsaI.rc site found at 2757