Part:BBa_K782078
10x[TALA+TALB] operator_CMV promoter_2xBFP:NLS
TALA and TALB labels represents TAL effector 1257 and 1297 from zebrafish experiments (Sander et al., 2011).
Introduction
Our construct contain TALA and TALB binding sites, which are adjacent to each other. Because of steric interference two different TAL regulators are unable to bind to its binding sites at once. TAL binding sites 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 730
- 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1406
Illegal BsaI.rc site found at 2062
Illegal BsaI.rc site found at 2767
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