Part:BBa_K747098:Design

MammoBrick

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 966
  Illegal suffix found in sequence at 1568
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 966
  Illegal SpeI site found at 1569
  Illegal PstI site found at 1583
  Illegal NotI site found at 972
  Illegal NotI site found at 1576
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 966
  Illegal BglII site found at 1563
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 966
  Illegal suffix found in sequence at 1569
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 966
  Illegal XbaI site found at 981
  Illegal SpeI site found at 1569
  Illegal PstI site found at 1583
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

See data in source

Source

BACKBONE:

the backbone is cut out of pTALEN (v2) NG It contains a SV 40 polyadenylation signal, an ampicillin resistance gene and an origin of replication.

CMV promoter:

see CMV-Promotor

PuroORF:

We replaced the hygromycin resistance gene in pTALEN (v2) NG with Puromycin. The gene is synthesized.

PostORF:

We called the region between the stop codon of the TAL ORF and the start codon of the antibiotic resistance gene PostORF. We wanted to use this part in our vector because it contains the SV40 promoter and enhancer for expression of the antibiotic selection marker

References

1. Scholze, H. & Boch, J. TAL effectors are remote controls for gene activation. Current Opinion in Microbiology 14, 47–53 (2011).

2. Moscou, M. J. & Bogdanove, A. J. A Simple Cipher Governs DNA Recognition by TAL Effectors. Science 326, 1501–1501 (2009).

3. Cermak, T. et al. Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res 39, e82 (2011).

4. Reyon, D. et al. FLASH assembly of TALENs for high-throughput genome editing. Nature Biotechnology 30, 460–465 (2012).

5. Zhang, F. et al. Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription. Nature biotechnology 29, 149–153 (2011).

6. Miller, J. C. et al. A TALE nuclease architecture for efficient genome editing. Nature Biotechnology 29, 143–148 (2010).

7. Boch, J. et al. Breaking the Code of DNA Binding Specificity of TAL-Type III Effectors. Science 326, 1509–1512 (2009).

8. Liu, J. et al. Efficient and Specific Modifications of the Drosophila Genome by Means of an Easy TALEN Strategy. Journal of Genetics and Genomics 39, 209–215 (2012).

9. Wood, A. J. et al. Targeted Genome Editing Across Species Using ZFNs and TALENs. Science 333, 307–307 (2011).

10. Sander, J. D. et al. Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nat Biotechnol 29, 697–698 (2011).

11. Tesson, L. et al. Knockout rats generated by embryo microinjection of TALENs. Nature Biotechnology 29, 695–696 (2011).

12. Hockemeyer, D. et al. Genetic engineering of human pluripotent cells using TALE nucleases. Nature Biotechnology 29, 731–734 (2011) 13. Sanjana, N. E. et al. A transcription activator-like effector toolbox for genome engineering. Nature Protocols 7, 171–192 (2012).