Difference between revisions of "Part:BBa K782035"
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<partinfo>BBa_K782035 short</partinfo> | <partinfo>BBa_K782035 short</partinfo> | ||
| − | TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011) | + | * TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011). |
| + | * DNA binding sites for individual TAL effectors are indicated with square brackets [ ]. | ||
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==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 [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782072 12] binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782005 TALD], upstream of a [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782088 minimal promoter]. Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter. | We designed our construct with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782072 12] binding sites for [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782005 TALD], upstream of a [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782088 minimal promoter]. Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter. | ||
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[[Image:12×D_pMIN_fLuc_slika1.png]] | [[Image:12×D_pMIN_fLuc_slika1.png]] | ||
| − | ''' | + | '''Figure 1: '''Schematic representation of our construct. |
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==Characterisation== | ==Characterisation== | ||
12x[TALD] operator_minimal promoter was characterised with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782031 mCitrine] reporter. | 12x[TALD] operator_minimal promoter was characterised with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782031 mCitrine] reporter. | ||
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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 22:11, 26 September 2012
12x[TALD] operator_minimal promoter_fLuciferase
- TALD label represents TAL effector 1295 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 12 binding sites for TALD, upstream of a minimal promoter. Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter.
Figure 1: Schematic representation of our construct.
Characterisation
12x[TALD] 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 749
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 847
Illegal NgoMIV site found at 2191
Illegal NgoMIV site found at 2212
Illegal NgoMIV site found at 2527
Illegal NgoMIV site found at 2635
Illegal AgeI site found at 460
Illegal AgeI site found at 692
Illegal AgeI site found at 1915 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 2097