Difference between revisions of "Part:BBa K782032"
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TALA and TALB labels represents TAL effectors 1257 and 1297 from zebrafish experiments (Sander et al., 2011)
 
TALA and TALB labels represents TAL effectors 1257 and 1297 from zebrafish experiments (Sander et al., 2011)
  
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===Introduction===
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==Introduction==
 
Transcription activation like (TAL) effectors are DNA binding proteins with a high specifity, built from tandem repeats, with  near identical seqences, differing only in two amino acids in each repeat called “repeat variable diresidue” (RVD), which determine the specifity for a single nucleotide.(Scholze and Boch, 2011)
 
Transcription activation like (TAL) effectors are DNA binding proteins with a high specifity, built from tandem repeats, with  near identical seqences, differing only in two amino acids in each repeat called “repeat variable diresidue” (RVD), which determine the specifity for a single nucleotide.(Scholze and Boch, 2011)
  
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'''Figure1:'''Schematic representation of our construct
 
'''Figure1:'''Schematic representation of our construct
  
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===Characterisation===
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==Characterisation==
  
 
10x[TALA+TALB] operator_minimal promoter was characterised with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782028 mCitrine] reporter.
 
10x[TALA+TALB] operator_minimal promoter was characterised with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782028 mCitrine] reporter.
  
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===References===
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==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
 
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

Revision as of 10:51, 26 September 2012

10x[TALA+TALB] operator_minimal promoter_fLuciferase TALA and TALB labels represents TAL effectors 1257 and 1297 from zebrafish experiments (Sander et al., 2011)

Introduction

Transcription activation like (TAL) effectors are DNA binding proteins with a high specifity, built from tandem repeats, with near identical seqences, differing only in two amino acids in each repeat called “repeat variable diresidue” (RVD), which determine the specifity for a single nucleotide.(Scholze and Boch, 2011)

We designed our construct with 10 binding sites for TALA and TALB, upstream of a minimal promoter (like here. Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter.

10×AB pMIN fLuc shema1.png

Figure1:Schematic representation of our construct

Characterisation

10x[TALA+TALB] operator_minimal promoter was characterised with mCitrine reporter.

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

Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1014
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 395
    Illegal NgoMIV site found at 760
    Illegal NgoMIV site found at 1112
    Illegal NgoMIV site found at 2456
    Illegal NgoMIV site found at 2477
    Illegal NgoMIV site found at 2792
    Illegal NgoMIV site found at 2900
    Illegal AgeI site found at 258
    Illegal AgeI site found at 598
    Illegal AgeI site found at 623
    Illegal AgeI site found at 963
    Illegal AgeI site found at 2180
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2362