Difference between revisions of "Part:BBa K782082"

 
 
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<partinfo>BBa_K782082 short</partinfo>
 
<partinfo>BBa_K782082 short</partinfo>
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* DNA binding sites for individual TAL effectors are indicated with square brackets [ ].
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==Introduction==
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Our construct contain twelve binding sites for [https://parts.igem.org/Part:BBa_K782007 NicTAL12] ([https://parts.igem.org/Part:BBa_K782003 binding site's sequence]) 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.
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[[Image:Nicbfp2.png‎‎]]
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'''Figure 1:''' Schematic representation of the construct.
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==Characterization==
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Results: Specific TAL [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782023 binding sites] were further characterized with other reporter constructs.
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BFP was obtained from Evrogen.
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==References==
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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.
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Latest revision as of 22:44, 26 September 2012

12x[NicTAL] operator_CMV promoter_2xBFP:NLS

  • DNA binding sites for individual TAL effectors are indicated with square brackets [ ].


Introduction

Our construct contain twelve binding sites for NicTAL12 (binding site's sequence) 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.

Nicbfp2.png

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 464
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 16
    Illegal AgeI site found at 248
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1139
    Illegal BsaI.rc site found at 1795
    Illegal BsaI.rc site found at 2500