Difference between revisions of "Part:BBa K782069"

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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 TALA.
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Our construct contain ten binding sites for TALA and another ten for TALB. Single binding site is combined from 18 base pairs recognised by specific TAL.TALA and TALB binding sites are adjacent to each other making impossible for both of them to bind at once.
 
   
 
   
Single binding site sequence for TALA: TTTACTGCTGCTCCCGCT
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Single binding site sequence for [https://parts.igem.org/Part:BBa_K782004 TALA]: 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.
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 +
Single binding site sequence for [https://parts.igem.org/Part:BBa_K782006 TALB]: TCTTCCGTTTCCACATCT
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[[Image:A.png]]
 
[[Image:A.png]]

Revision as of 16:50, 25 September 2012

10x[TALA+TALB] operator


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 for TALA and another ten for TALB. Single binding site is combined from 18 base pairs recognised by specific TAL.TALA and TALB binding sites are adjacent to each other making impossible for both of them to bind at once.

Single binding site sequence for TALA: TTTACTGCTGCTCCCGCT

Single binding site sequence for TALB: TCTTCCGTTTCCACATCT


A.png

Figure 1: Schematic representation of the construct.


Characterization

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



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 149
    Illegal NgoMIV site found at 514
    Illegal AgeI site found at 12
    Illegal AgeI site found at 352
    Illegal AgeI site found at 377
    Illegal AgeI site found at 717
  • 1000
    COMPATIBLE WITH RFC[1000]