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(Construction details)
 
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=== Construction details ===
 
=== Construction details ===
pME-EGFP was made by PCR amplification of EGFP, using primers that add att sites, followed by a BP reaction.  
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pME-EGFP was made by PCR amplification of the EGFP insert from a pCS2-based plasmid, using primers that add att sites, followed by a BP reaction. Sequencing confirms that all bases of the insert are correct.
Sequencing confirms that all bases of the insert are correct.
 
  
 
Note that the EGFP sequence is followed by 60 bp of polylinker sequence from pCS2+ before the stop codon. We have confirmed functionally that this cassette produces fluorescent protein.
 
Note that the EGFP sequence is followed by 60 bp of polylinker sequence from pCS2+ before the stop codon. We have confirmed functionally that this cassette produces fluorescent protein.
  
=== Crudely annotated sequence ===
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=== Sequence ===
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Annotated sequence, Genbank format:<br>
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[[pME-EGFP Genbank]]
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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>
 
[[pME-EGFP sequence]]
 
[[pME-EGFP sequence]]

Latest revision as of 03:44, 19 December 2007

Construction details

pME-EGFP was made by PCR amplification of the EGFP insert from a pCS2-based plasmid, using primers that add att sites, followed by a BP reaction. Sequencing confirms that all bases of the insert are correct.

Note that the EGFP sequence is followed by 60 bp of polylinker sequence from pCS2+ before the stop codon. We have confirmed functionally that this cassette produces fluorescent protein.

Sequence

Annotated sequence, Genbank format:
pME-EGFP Genbank

FASTA file with the full-length sequence as well as sequences of individual components:
pME-EGFP sequence

Crude map

Screenshot from Sequencher showing locations of components:
PME-EGFP.png