Addgene - LHP1263 - HA-VPS9 Plasmid Data

Plasmid 32431: LHP1263 - HA-VPS9

Gene/insert name:
VPS9
Species:
S. cerevisiae (budding yeast)
Entrez Gene:
VPS9 (YML097C, VPL31, VPT9)
Fusion protein or tag:
3xHA
Terminal:
N terminal on insert
Vector backbone:
YCplac111
(Search Vector Database)
Vector type:
Yeast Expression
Promoter:
Lac
Cloning site 5':
none
Site destroyed during cloning:
Unknown
Cloning site 3':
none
Site destroyed during cloning:
Unknown
5' sequencing primer:
M13 reverse List of Sequencing Primers
3' sequencing primer:
M13 forward
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Selectable markers:
LEU2
Sequence: View sequences (1)
Principal Investigator:
Linda Hicke
Terms and Licenses:
MTA

Comments:

To insert an HA epitope at the N-terminus of Vps9, a NotI site was introduced after the first methionine codon of VPS9 in pPS91 (Hama et al., 1999). NotI-VPS9 was then subcloned into the centromere-based vector YCplac111 and a fragment encoding an 3xHA tag flanked by NotI sites was inserted.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: A ubiquitin-binding motif required for intramolecular monoubiquitylation, the CUE domain. Shih et al (EMBO J. 2003 Mar 17;22(6):1273-81. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 32431" in your Materials and Methods section.