Difference between revisions of "Part:BBa K747000"

 
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<partinfo>BBa_K747000 short</partinfo>
 
<partinfo>BBa_K747000 short</partinfo>
  
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This Part specifies the DNA binding affinity of the 2. and 3. base pair of a Transactivator-like (TAL) protein. The TAL-Protein binds a specific fourteen base pair DNA sequence. The 2. and 3. base pair of this part is specified by adenine-adenine.
 
  
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'''TAL- Proteins'''
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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.
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'''Designing a TAL-Protein:'''
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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.
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Bba_K747016 to  Bba_K747031 predefines the nucleotide of the 4. and the 5. nucleotide.
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Bba_K747032 to  Bba_K747047 predefines the nucleotide of the 6. and the 7. nucleotide.
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Bba_K747048 to  Bba_K747063 predefines the nucleotide of the 8. and the 9. nucleotide.
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Bba_K747064 to  Bba_K747079 predefines the nucleotide of the 10. and the 11. nucleotide.
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Bba_K747080 to  Bba_K747095 predefines the nucleotide of the 12. and the 13. nucleotide.
  
 
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===Usage and Biology===
 
===Usage and Biology===
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'''Constructing a TAL-Protein:'''
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Only one Golden Gate Cloning step is necessary to insert the Direpeats into a appropriate vector e.g.  Bba_K747096 to  Bba_K747098. Every DiRepeat is flanked by a BsmBI restriction site which determines its location in the TAL protein.
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Use a vector:insert ratio of 1:1 for the ligation. Following protocol should be used:
  
 
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Revision as of 16:17, 24 September 2012

TAL-Protein_AA1_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_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.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 166