pENTR-FKBP12DD Vectors

Example of FKBP-DD-fused Asf1a:

 

The Asf1a cDNA was inserted into the pENTR4-FKBP12DD N-term. C (w385-10) plasmid (see below) and recombined into the pLenti CMV Hygro DEST (w117-1) vector. Viral particles were used to transduce HeLa cells and transduced cells were selected with Hygromycin. When the cells are brown in the presence of Shld-1, the FKBP-Asf1a fusion protein can be detected (lane 1); upon removal of Shld-1, the FKBP-Asf1a fusion protein can no longer be detected (lane 2). Endogenous levels of Asf1a are unaffected by the presence of the FKBP-As1fa fusion protein. Alpha-tubulin is used as a loading control. The FKBP-Asf1a can also be degraded in cells arrested in mitosis (Campeau et al. 2009).

 

pENTR4-FKBP12DD N-term. A (B3-1)

High-resolution pdf: pENTR-FKBPDD N-term A (B3-1).pdf

Sequence file: pENTR-FKBPDD N-term A (B3-1).seq

Sequence verification file: B3-1ENTRforw.ab1

Addgene #:17414

Recommended E.coli strain for propagation: TOP10F'.

Selection: Kanamycin.

Recommended Destination vectors: pLenti CMV, CMV/TO, PGK series, pQCX CMV/TO series

 

pENTR4-FKBP12DD N-term. B (w385-4)

High-resolution pdf: pENTR-FKBP12DD N-term B (w385-4).pdf

Sequence file: pENTR-FKBP12DD N-term B (w385-4).seq

Sequence verification file: w385-4ENTRrev.ab1

Addgene #:17415

Recommended E.coli strain for propagation: TOP10F'.

Selection: Kanamycin.

Recommended Destination vectors: pLenti CMV, CMV/TO, PGK series, pQCX CMV/TO series

 

pENTR4-FKBP12DD N-term. C (w385-10)

High-resolution pdf: pENTR-FKBP12DD N-term C (w385-10).pdf

Sequence file: pENTR-FKBP12DD N-term C (w385-10).seq

Sequence verification file: w385-10ENTRrev.ab1

Addgene #:17416

Recommended E.coli strain for propagation: TOP10F'.

Selection: Kanamycin.

Recommended Destination vectors: pLenti CMV, CMV/TO, PGK series, pQCX CMV/TO series

 

pENTR1A-FKBP12DD C-term. (w350-2)

High-resolution pdf: pENTR-FKBP12DD C-term (w350-2).pdf

Sequence file: pENTR-FKBP12DD C-term (w350-2).seq

Sequence verification file: w350-2ENTRforw.ab1

Addgene #:17413

Recommended E.coli strain for propagation: TOP10F'.

Selection: Kanamycin.

Recommended Destination vectors: pLenti CMV, CMV/TO, PGK series, pQCX CMV/TO series