From Tol2Kit
Jump to: navigation, search
(Construction details)
(Construction details)
Line 1: Line 1:
 
=== Construction details ===
 
=== Construction details ===
 
pME-EGFP was made by PCR amplification of EGFP, using primers that add att sites, followed by a BP reaction.  
 
pME-EGFP was made by PCR amplification of EGFP, 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.
+
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.
  
 
=== Crudely annotated sequence ===
 
=== Crudely annotated sequence ===

Revision as of 01:39, 24 March 2007

Construction details

pME-EGFP was made by PCR amplification of EGFP, 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.

Crudely annotated sequence

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