Protein_Domain

Part:BBa_K747002

Designed by: Lucas Schneider   Group: iGEM12_Freiburg   (2012-09-24)
Revision as of 14:06, 25 September 2012 by Lucas (Talk | contribs)

TAL-Protein_AG1_Direpeat

TAL- Proteins This part can be used to build a 14 nucleotide Transactivator-like (TAL) protein. TAL- Proteins are DNA-targeting modules with repeating Domains, each one of them can bind a specific nucleotide. A plasmid set of 96 DiRepeats (Bba_K747000 to Bba_K747095) allows you to assemble your own specific nucleotide target sequence. Every DiRepeat is able to bind two specific nucleotides and is determined by his position in the 14 repeating Domains.

Designing a TAL-Protein: To build a functional Tal- Protein, you have to choose a 14 nucleotide target sequence. Be aware that the first and the fourteenth nucleotide of the sequence has to be a Thymin. The second to the thirteenth nucleotide can be determined by choosing six DiRepeats of the plasmid set: Bba_K747000 to Bba_K747015 predefines the nucleotide of the 2. and the 3. nucleotide.

  • Bba_K747000 to Bba_K747015 predefines the nucleotide of the 2. and the 3. nucleotide.
  • Bba_K747016 to Bba_K747031 predefines the nucleotide of the 4. and the 5. nucleotide.
  • Bba_K747032 to Bba_K747047 predefines the nucleotide of the 6. and the 7. nucleotide.
  • Bba_K747048 to Bba_K747063 predefines the nucleotide of the 8. and the 9. nucleotide.
  • Bba_K747064 to Bba_K747079 predefines the nucleotide of the 10. and the 11. nucleotide.
  • Bba_K747080 to Bba_K747095 predefines the nucleotide of the 12. and the 13. nucleotide.

Pipette the following volumes in one tube:

Component Volume
6 Direpeats of your choice 63,3 ng
Backbone e. g. TAL-TF 200 ng ???
BsmBI 1,5 µl
T4 DNA Ligase (60 u) 1,0 µl
T4 DNA Ligase Buffer 2,0 µl
Water, nuclease-free up to 20 µl
Total vloume 20 µl




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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//proteindomain/dnabinding
Parameters
None