P5E-MCS

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[edit] Construction details

p5E-MCS was made by PCR of the pBSII SK+ multiple cloning site, from T7 to T3 primers, followed by a BP reaction with pDONR P4-P1R. Sequencing confirms that all bases of the insert are correct.

M13F, M13R, and T3 should all work as sequencing primers.

Unfortunately, the backbone of P4-P1R contains recognition sites for several potentially useful restriction enzymes. The restriction sites from the Bluescript MCS that remain unique in p5E-MCS are:

Asp718-KpnI-DraII-XhoI-SalI-Bsp106-HindIII-SmaI-BamHI-SpeI-SacII-BstXI

[edit] Sequence

Annotated sequence, Genbank format:
p5E-MCS Genbank

FASTA file with the full-length sequence as well as sequences of individual components:
p5E-MCS sequence

[edit] Crude map

Screenshot from Sequencher showing locations of components:
Image:p5E-MCS.png

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