Part:BBa_K782088

10x[TALB] operator_minimal promoter

• TALB and TALC labels represents TAL effector 1297 and 1259 from zebrafish experiments (Sander et al., 2011).

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 TALB binding sites and minimal promoter.

Minimal promotor is taken from pGL4 vector series([http://www.promega.com/~/media/files/resources/protocols/technical%20manuals/0/pgl4%20luciferase%20reporter%20vectors%20protocol.pdf?la=en Ref] promotor is described on the page 18)

Single binding site is combined from 18 base pairs specificly recognised by TALB. Binding sites are constructed in alternating setup. Besides TALB:VP16 binding sites construct also contain ten alternating TALC:VP16 binding sites, which we didn't use, since TALC:VP16 regulator wasn't working properly.

Single binding site sequence for TALB: TCTTCCGTTTCCACATCT

Figure 1: Schematic representation of the construct.

Characterization

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 394
  Illegal NgoMIV site found at 754
  Illegal AgeI site found at 259
  Illegal AgeI site found at 594
  Illegal AgeI site found at 619
  Illegal AgeI site found at 954
  
• 1000
  COMPATIBLE WITH RFC[1000]