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Revision as of 23:32, 27 September 2007 by Chi-bin (talk | contribs) (Construction details)

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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, and the 3' end of the M13R sequence, M13F will definitely not work for sequencing clones made from pME-MCS (it will bind both in the vector backbone and in the insert), and M13R will work poorly (it will bind perfectly to the vector backbone and weakly in the insert).

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