Difference between revisions of "Part:BBa K782073"
 
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<partinfo>BBa_K782073 short</partinfo>
 
<partinfo>BBa_K782073 short</partinfo>
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* TALA and TALB labels represents TAL effector 1257 and 1297 from zebrafish experiments (Sander et al., 2011).
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* DNA binding sites for individual TAL effectors are indicated with square brackets [ ].
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==Introduction==
 
==Introduction==
  
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Our construct contain [https://parts.igem.org/Part:BBa_K782069 TALA and TALB binding sites], which are adjacent to each. For that reason two different TAL regulators are unable to bind to its binding sites at once because of steric interference. TAL binding sites are cloned downstream of CMV promoter. Upstream of CMV is Blue fluorescent protein. BFP in a monomeric fluorescent protein with excitation maximum at 402 nm and emision maximum at 457 nm.
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Our construct contain [https://parts.igem.org/Part:BBa_K782069 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 blue fluorescent protein. BFP is a monomeric fluorescent protein with excitation maximum at 402 nm and emision maximum at 457 nm.
  
 
[[Image:Abbfp.png‎]]
 
[[Image:Abbfp.png‎]]
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Results:  
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Results: Specific TAL [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782019 binding sites] were further characterized with other reporter constructs.
  
 
BFP was obtained from Evrogen.
 
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.
  
  

Latest revision as of 22:33, 26 September 2012

10x[TALA+TALB] operator_CMV promoter_BFP

  • TALA and TALB labels represents TAL effector 1257 and 1297 from zebrafish experiments (Sander et al., 2011).
  • DNA binding sites for individual TAL effectors are indicated with square brackets [ ].


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 blue fluorescent protein. BFP is a monomeric fluorescent protein with excitation maximum at 402 nm and emision maximum at 457 nm.

Abbfp.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 730
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1406
    Illegal BsaI.rc site found at 2062