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Vectors

pLPPI-SIRT7-FLAG-STrEPII (click here for sequence) – This is a lentiviral vector using the pLL3.7 backbone.  All the non-viral elements were cut out of pLL3.7 using XbaI and PciI.  Inserted in the following order was the PGK promoter, the puromycin resistance gene, IRES, SIRT7, FLAG tag, STrEP II tag and a stop codon.  To clone in other genes of interest, use the XhoI site and the NheI site.  The reading frame splits down the middle of the NheI site.  The PGK promoter is nice because it is constitutive but expresses at low levels in many cell types.  This allows for closer to endogenous levels of expression.  The STrEP tag is great for large scale purifications and the FLAG tag is good tag for smaller scale purifications.  The two work well together for tandem affinity purification (TAP).

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