Difference between revisions of "Part:BBa K3338011"

 
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===Usage and Biology===
  
 
The composite part described here exhibits an EGFP-MagA-P2A-hGLuc-cassette under the control of a CMV-enhancer/promoter for expression in mammalian cells. It was used to determine the expression of MagA and hGLuc at the same time from one promoter.
 
The composite part described here exhibits an EGFP-MagA-P2A-hGLuc-cassette under the control of a CMV-enhancer/promoter for expression in mammalian cells. It was used to determine the expression of MagA and hGLuc at the same time from one promoter.
  
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=Cloning=
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===Usage and Biology===
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===Theoretical Part Design===
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In order to test whether HeLa cells are able to simultaneously express MagA and hGLuc using a P2A-peptide, MagA and hGLuc were cloned into the pEGFP-C2 vector interspaced with P2A. In the resulting plasmid MagA is encoded with a N-terminal EGFP-tag making MagA detection very easy. The CMV promoter ensures constitutive and high expression rates mimicking the fully activated sensor.
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===Sequence and Features===
  
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<span class='h3bb'>Sequence and Features</span>
 
 
<partinfo>BBa_K3338011 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3338011 SequenceAndFeatures</partinfo>
  
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===Cloning===
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pEGFP-C2 was cut with
  
 
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Revision as of 14:03, 23 October 2020


CMV-EGFP-MagA-P2A-hGLuc

Usage and Biology

The composite part described here exhibits an EGFP-MagA-P2A-hGLuc-cassette under the control of a CMV-enhancer/promoter for expression in mammalian cells. It was used to determine the expression of MagA and hGLuc at the same time from one promoter.

Cloning

Theoretical Part Design

In order to test whether HeLa cells are able to simultaneously express MagA and hGLuc using a P2A-peptide, MagA and hGLuc were cloned into the pEGFP-C2 vector interspaced with P2A. In the resulting plasmid MagA is encoded with a N-terminal EGFP-tag making MagA detection very easy. The CMV promoter ensures constitutive and high expression rates mimicking the fully activated sensor.

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 site found at 2553
    Illegal BsaI.rc site found at 1885
    Illegal BsaI.rc site found at 2434
    Illegal SapI site found at 1575


Cloning

pEGFP-C2 was cut with