Difference between revisions of "Part:BBa K782070"
 
(11 intermediate revisions by 3 users not shown)
Line 2: Line 2:
 
<partinfo>BBa_K782070 short</partinfo>
 
<partinfo>BBa_K782070 short</partinfo>
  
−
 
+
* TALA and TALC labels represents TAL effector 1257 and 1259 from zebrafish experiments (Sander et al., 2011).
 +
* DNA binding sites for individual TAL effectors are indicated with square brackets [ ].
  
 
==Introduction==
 
==Introduction==
Line 8: Line 9:
 
TAL effectors (TALEs) are able to specifically bind to the DNA by recognizing base pairs on a DNA molecule. With TAL regulators we are able to control expression of a desired gene. Efficient transcription control can be achieved by binding of multiple TAL regulators upstream of promoter.  
 
TAL effectors (TALEs) are able to specifically bind to the DNA by recognizing base pairs on a DNA molecule. With TAL regulators we are able to control expression of a desired gene. Efficient transcription control can be achieved by binding of multiple TAL regulators upstream of promoter.  
  
−
Construct contain ten binding sites while single binding site combines 18 base pairs specificly recognised by [https://parts.igem.org/Part:BBa_K782004 TALA].
+
Our construct contain ten binding sites while single binding site is combined from 18 base pairs specificly recognised by TALA . Binding sites are constructed in alternating setup.  
 +
 
 +
Besides TALA binding sites construct also contain ten alternating TALC binding sites, which we didn't use, since TALC regulator wasn't working properly.
 
   
 
   
−
Single binding site sequence for TALA: TTTACTGCTGCTCCCGCT
+
Single binding site sequence for [https://parts.igem.org/Part:BBa_K782004 TALA] is: TTTACTGCTGCTCCCGCT
−
Binding sites are constructed in alternating setup. Besides TALA binding sites construct also contain ten alternating TALC binding sites, which we didn't use, since TALC wasn't working properly.
+
  
−
[[Image:A.png]]
+
 
 +
 
 +
[[Image:10xA.png]]
  
 
'''Figure 1:''' Schematic representation of the construct.
 
'''Figure 1:''' Schematic representation of the construct.
 +
 +
 +
 +
* Binding sites for TAL effectors were ordered from GeneArt.
  
  
 
==Characterization==
 
==Characterization==
  
−
Construct was later used for constructing different reporter plasmids to show specific binding of TALA KRAB / VP16.
+
Construct was later used for constructing different reporter plasmids to show specific binding of [https://parts.igem.org/Part:BBa_K782008 TALA:KRAB]/[https://parts.igem.org/Part:BBa_K782065 VP16].
−
Results:  
+
 
 +
'''Results:'''
 +
 
 +
* [https://parts.igem.org/Part:BBa_K782017 BBa_K782017]
  
  
 +
==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.
  
  

Latest revision as of 22:54, 26 September 2012

10x[TALA] operator

  • TALA and TALC labels represents TAL effector 1257 and 1259 from zebrafish experiments (Sander et al., 2011).
  • DNA binding sites for individual TAL effectors are indicated with square brackets [ ].

Introduction

TAL effectors (TALEs) are able to specifically bind to the DNA by recognizing base pairs on a DNA molecule. With TAL regulators we are able to control expression of a desired gene. Efficient transcription control can be achieved by binding of multiple TAL regulators upstream of promoter.

Our construct contain ten binding sites while single binding site is combined from 18 base pairs specificly recognised by TALA . Binding sites are constructed in alternating setup.

Besides TALA binding sites construct also contain ten alternating TALC binding sites, which we didn't use, since TALC regulator wasn't working properly.

Single binding site sequence for TALA is: TTTACTGCTGCTCCCGCT


10xA.png

Figure 1: Schematic representation of the construct.


  • Binding sites for TAL effectors were ordered from GeneArt.


Characterization

Construct was later used for constructing different reporter plasmids to show specific binding of TALA:KRAB/VP16.

Results:


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.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 147
    Illegal NgoMIV site found at 507
    Illegal AgeI site found at 12
    Illegal AgeI site found at 347
    Illegal AgeI site found at 372
    Illegal AgeI site found at 707
  • 1000
    COMPATIBLE WITH RFC[1000]