Difference between revisions of "Part:BBa K782032"
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<partinfo>BBa_K782032 short</partinfo> | <partinfo>BBa_K782032 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 [ ]. | ||
| + | |||
==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] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782006 TALB], upstream of a minimal promoter (like [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782086 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] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782006 TALB], upstream of a minimal promoter (like [https://parts.igem.org/wiki/index.php?title=Part:BBa_K782086 here]). Downstream of the promoter we cloned fLuciferase (firefly luciferase) to function as a reporter. |
[[Image:10×AB_pMIN_fLuc_shema1.png]] | [[Image:10×AB_pMIN_fLuc_shema1.png]] | ||
| − | ''' | + | '''Figure 1:'''Schematic representation of our construct. |
| + | |||
==Characterisation== | ==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. | ||
| + | |||
==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 22:08, 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).
- 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 and TALB, 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+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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1014
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 2362