Difference between revisions of "Part:BBa K1061011"
 
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Ef-1alpha promoter is a strong constitutive promoter  that drive the expression of elongation factor -1α.It has been widely used in constructing mammalian cell-based circuit, especially useful for construction of long-term circuit in some cells including iPS cells and Jurkat cells in which the strong promoter CMV may be silenced.[1]
 
Ef-1alpha promoter is a strong constitutive promoter  that drive the expression of elongation factor -1α.It has been widely used in constructing mammalian cell-based circuit, especially useful for construction of long-term circuit in some cells including iPS cells and Jurkat cells in which the strong promoter CMV may be silenced.[1]
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===Further Characterization - Team Technion 2017===
 
===Further Characterization - Team Technion 2017===
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Team Technion 2017 created a mutated version of the full EF-1alpha promoter <partinfo>K2520023</partinfo>.  
 
Team Technion 2017 created a mutated version of the full EF-1alpha promoter <partinfo>K2520023</partinfo>.  
 
This promoter is fully functional and lacks the forbidden restriction sites.  
 
This promoter is fully functional and lacks the forbidden restriction sites.  
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===Usage and Biology===
 
===Usage and Biology===
  

Latest revision as of 19:38, 31 October 2017

EF-1alpha promoter

Ef-1alpha promoter is a strong constitutive promoter that drive the expression of elongation factor -1α.It has been widely used in constructing mammalian cell-based circuit, especially useful for construction of long-term circuit in some cells including iPS cells and Jurkat cells in which the strong promoter CMV may be silenced.[1]

Further Characterization - Team Technion 2017

We believe this is not the full EF-1alpha promoter, but rather the core promoter of EF-1alpha (Approx. 300bp) attached to the R segment and part of the U5 sequence (R-U5’) of the Human T-Cell Leukemia Virus (HTLV) Type 1 Long Terminal Repeat.

The full EF-1alpha promoter is approx. 1200bp, and contains two forbidden restriction sites (by iGEM Biobrick standards).

Team Technion 2017 created a mutated version of the full EF-1alpha promoter BBa_K2520023. This promoter is fully functional and lacks the forbidden restriction sites.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 344
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 440
  • 1000
    COMPATIBLE WITH RFC[1000]


References:

[1] R.V. Gopalkrishnan, K.A. Christiansen et al, 1999,Use of the human EF-1α promoter for expression can significantly increase success in establishing stable cell lines with consistent expression: A study using the tetracycline-inducible system in human cancer cells, nucleic acids research,27:4775-4782