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'''Construction details'''<br>
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=== Construction details ===
p3E-IRES-nlsEGFPpA was made by fusion PCR of the EMCV IRES sequence, and nlsEGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of the coding sequence are correct. There is a single base change in the IRES sequence, but we have confirmed functionally that this cassette produces fluorescent, nuclear-localized protein.
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p3E-IRES-nlsEGFPpA was made by fusion PCR of the EMCV IRES sequence (IRES2 from Clontech), and nlsEGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of the coding sequence are correct. There is a single base change in the IRES sequence (G481A), but we have confirmed functionally that this cassette produces fluorescent, nuclear-localized protein.
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=== Sequence ===
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Annotated sequence, Genbank format:<br>
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[[p3E-IRES-nlsEGFPpA Genbank]]
  
'''Crudely annotated sequence'''<br>
 
 
FASTA file with the full-length sequence as well as sequences of individual components:<br>
 
FASTA file with the full-length sequence as well as sequences of individual components:<br>
 
[[p3E-IRES-nlsEGFPpA sequence]]
 
[[p3E-IRES-nlsEGFPpA sequence]]
  
'''Crude map'''<br>
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=== Crude map ===
 
Screenshot from Sequencher showing locations of components:<br>
 
Screenshot from Sequencher showing locations of components:<br>
 
[[Image:p3E-IRES-nlsEGFPpA.png]]
 
[[Image:p3E-IRES-nlsEGFPpA.png]]

Latest revision as of 18:47, 17 August 2011

Construction details

p3E-IRES-nlsEGFPpA was made by fusion PCR of the EMCV IRES sequence (IRES2 from Clontech), and nlsEGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of the coding sequence are correct. There is a single base change in the IRES sequence (G481A), but we have confirmed functionally that this cassette produces fluorescent, nuclear-localized protein.

Sequence

Annotated sequence, Genbank format:
p3E-IRES-nlsEGFPpA Genbank

FASTA file with the full-length sequence as well as sequences of individual components:
p3E-IRES-nlsEGFPpA sequence

Crude map

Screenshot from Sequencher showing locations of components:
P3E-IRES-nlsEGFPpA.png