Difference between revisions of "Part:BBa K782036"
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+ | TALB, TALA and TALD labels represents TAL effectors 1297 and 1257 and 1295 respectively from zebrafish experiments (Sander et al., 2011) | ||
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+ | ===Introduction=== | ||
+ | TAL effectors (TALEs) are bacterial plant pathogen transcription factors, that bind to DNA by specifically recognizing one base pair with a single tandem repeat in their DNA-binding domain. A tandem TALE repeat contains 33 to 35 amino acids, where the 12th and 13th amino acid, called a “repeat variable diresidue” (RVD), are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011). | ||
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+ | This part contains 10 alterations of TALA and TALB binding sites ([https://parts.igem.org/Part:BBa_K782069 10×[TALA+TALB]]), upstream of a CMV promoter (constitutive promoter for expression in mammalian cells). Downstream of the promoter there is a TAL repressor [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782009 TALD:NLS:KRAB]. This way we have a TAL effector under the control of other TAL effectors, which allows us to create more complicated genetic circuits with more than one level of regulation. | ||
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+ | [[Image:SVN12_10×AB_TDK.png]] | ||
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+ | '''Figure 1:'''Schematic representation of the construct | ||
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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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+ | Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53 | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 17:56, 25 September 2012
10x[TALA+TALB] operator_CMV promoter_TALD:NLS:KRAB
TALB, TALA and TALD labels represents TAL effectors 1297 and 1257 and 1295 respectively from zebrafish experiments (Sander et al., 2011)
Introduction
TAL effectors (TALEs) are bacterial plant pathogen transcription factors, that bind to DNA by specifically recognizing one base pair with a single tandem repeat in their DNA-binding domain. A tandem TALE repeat contains 33 to 35 amino acids, where the 12th and 13th amino acid, called a “repeat variable diresidue” (RVD), are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011).
This part contains 10 alterations of TALA and TALB binding sites (10×[TALA+TALB]), upstream of a CMV promoter (constitutive promoter for expression in mammalian cells). Downstream of the promoter there is a TAL repressor TALD:NLS:KRAB. This way we have a TAL effector under the control of other TAL effectors, which allows us to create more complicated genetic circuits with more than one level of regulation.
Figure 1:Schematic representation of the construct
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 924
Illegal BamHI site found at 1556
Illegal BamHI site found at 3924 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 343
Illegal NgoMIV site found at 708
Illegal AgeI site found at 206
Illegal AgeI site found at 546
Illegal AgeI site found at 571
Illegal AgeI site found at 911 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 4290
Illegal SapI.rc site found at 4254