Difference between revisions of "Part:BBa K782033"
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<partinfo>BBa_K782033 short</partinfo> | <partinfo>BBa_K782033 short</partinfo> | ||
| − | * 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). |
* DNA binding sites for individual TAL effectors are indicated with square brackets [ ]. | * DNA binding sites for individual TAL effectors are indicated with square brackets [ ]. | ||
==Introduction== | ==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). |
We designed our construct with 10 binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782004 TALA], upstream of a minimal promoter (like [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782087 here]). Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter. | We designed our construct with 10 binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782004 TALA], upstream of a minimal promoter (like [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782087 here]). Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter. | ||
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[[Image:10×A_pMIN_fLuc_shema1.png]] | [[Image:10×A_pMIN_fLuc_shema1.png]] | ||
| − | ''' | + | '''Figure 1:'''Schematic representation of our construct. |
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==References== | ==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. |
| − | Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53 | + | Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53. |
Latest revision as of 22:09, 26 September 2012
10x[TALA] operator_minimal promoter_fLuciferase
- TALA and TALB labels represents TAL effectors 1257 and 1297 from zebrafish experiments (Sander et al., 2011).
- DNA binding sites for individual TAL effectors are indicated with square brackets [ ].
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, upstream of a minimal promoter (like here). Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter.
Figure 1:Schematic representation of our construct.
Characterisation
10x[TALA] 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1004
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 393
Illegal NgoMIV site found at 753
Illegal NgoMIV site found at 1102
Illegal NgoMIV site found at 2446
Illegal NgoMIV site found at 2467
Illegal NgoMIV site found at 2782
Illegal NgoMIV site found at 2890
Illegal AgeI site found at 258
Illegal AgeI site found at 593
Illegal AgeI site found at 618
Illegal AgeI site found at 953
Illegal AgeI site found at 2170 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 2352