Part:BBa_K1789007
SCAF3
This part is a scaffold system contains the recognizing motifs of TALE1 and TALE2
Usage and Biology
As is mentioned in our project description, different TALEs share a similar domain structure that enables them to bind the genome of the host cell and act as transcriptional effectors. By engineering those structures, we can build proteins that can bind with any DNA sequence that we desire.
In our project, we designed two different DNA binding motifs (DNA BMs). The sequences were chosen from Danio rerio CD154 gene in order to avoid homology with E.coli genome. Those BMs are sequenced as
BM1: 5’-GGAGGCACCGGTGG-3’
MB2: 5’-GATAAACACCTTTC-3’
Those sequences were repeated for more than 10 times in a plasmid, with different length of intervening sequence.
This scaffold is designed to put enzymes in the same direction. A 16-bp intervening sequence is added between two BMs.
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 8
Illegal AgeI site found at 70
Illegal AgeI site found at 132
Illegal AgeI site found at 194
Illegal AgeI site found at 256
Illegal AgeI site found at 318
Illegal AgeI site found at 380 - 1000COMPATIBLE WITH RFC[1000]
Experimental Validation
Sequencing
This part is sequenced as correct after construction.
Enzyme digestion test
A double-enzyme digestion test was implemented to verify this part.
The length of the digested DNA fits the design.
Functional Test
The function of this part can be verified with in the experimental validation of BBa_K1789022.
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