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Revision as of 23:30, 27 September 2007 by Chi-bin (talk | contribs) (New page: === Construction details === pME-MCS was made by PCR of the pBSII SK+ multiple cloning site, from M13F to M13R primers, followed by a BP reaction with pDONR221. Sequencing confirms that al...)

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Construction details

pME-MCS was made by PCR of the pBSII SK+ multiple cloning site, from M13F to M13R primers, followed by a BP reaction with pDONR221. Sequencing confirms that all bases of the insert are correct.

Since the insert contains the entire M13F sequence, but only part of the M13R sequence, M13F will not be useful for sequencing clones made from pME-MCS.

The following sites from the Bluescript MCS remain unique in pME-MCS:

KpnI-XhoI-SalI-Bsp106-HindIII-EcoRI-PstI-SmaI-BamHI-SpeI-XbaI-NotI-XmaIII-SacII-BstXI-SacI

Crudely annotated sequence

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

Crude map

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