Part:BBa_K782070
10x[TALA] operator
TALA and TALC labels represents TAL effector 1257 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.
Construct contain ten binding sites while single binding site combines 18 base pairs specificly recognised by TALA.
Single binding site sequence for 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 regulator wasn't working properly.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]
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