pYDL-12

pYDL-12

Technical Description

pYDL™-12 is a vector designed for yeast display of target proteins fused to the N-terminus of Aga2p for anchoring to the cell wall of Aga1p-expressing tryptophan-auxotrophic S. cerevisiae (i.e., EBY100, or equivalent) (Fig. 1). The vector contains the appS4 leader sequence followed by a double BsmBI cloning site to introduce foreign polypeptides on the N-terminal side of the Aga2p protein with a HA-tag for detection. Expression of the fusion is under the tight control of the GAL1 promoter, allowing for strong metabolic induction with galactose in the absence of glucose.

Figure 1. pYDL-12 Surface Display and Binding Analysis by Flow Cytometry. Schematic representation of pYDL-12 display (Left Panel). Flow cytometry analysis of single-domain antibody display (FITC-A) and target binding (APC-A) by clonal yeast transformed with pYDL-12 (Right Panel).

Applications

• Yeast display at the N-terminal side of Aga2p protein.

• Display of single domain antibodies, scFv, peptides or similar scaffolds.

For research use only; not intended for any animal or human therapeutic or diagnostic use.

Specifications

General Characteristics:

Plasmid Size: 4514
Promoter: GAL1 promoter
Leader Peptide: appS4
Cloning Site: double-BsmBI
Fusion Protein: full length Aga2p protein with HA-tag
Selection: ampicillin (bacterial), tryptophan deficiency (yeast)
Replication: pMB1 (bacterial, high copy number), CEN/ARS (yeast, low copy number)

Physical Characteristics:

Concentration: 0.5 µg/µl.
Product Size: 10 µg.
Buffer: DNA Conservation Buffer (Tris/HCL 5 mM, EDTA 0.1 mM, pH 8.5, sterile).
Storage Temperature: -20°C.

Quality Control & Certification of Analysis

Product Sequence:

Complete vector sequence is analyzed.

Certification:

Product meets all specifications.

Related Products

pYDL Yeast Display Vector Series