pKLAC2 Vector

Catalog # N3742 was discontinued on January 02, 2018 ;

The pKLAC2 vector directs high-level expression of a recombinant protein from the yeast Kluyveromyces lactis and is compatible with the K. lactis Protein Expression Kit (NEB #E1000). pKLAC2 can be used for either intracellular or secreted protein expression. SacII or BstXI linearized pKLAC2 integrates into the LAC4 locus of the K. lactis genome upon transformation of K. lactis competent cells.

Contains a universal multiple cloning site that is compatible with all NEB expression systems
PLAC4-PBI promoter does not express in E. coli, thereby allowing toxic genes to be cloned prior to their expression in yeast
Includes acetamidase expression for non-antibiotic selection in K. lactis
Includes ampicillin resistance for propagation in E. coli

Product Information
The vector pKLAC2 directs high-level expression of a recombinant protein from the yeast Kluyveromyces lactis and is compatible with the K. lactis Protein Expression Kit (NEB #E1000). pKLAC2 can be used for either intracellular or secreted protein expression. SacII or BstXI linearized pKLAC2 integrates into the LAC4 locus of the K. lactis genome upon transformation of K. lactis competent cells. pKLAC2 differs from pKLAC1 (>NEB #N3740) in that it contains a universal multiple cloning site (MCS) that is compatible with all NEB expression systems.

Vector pKLAC2 contains the strong K. lactis P_LAC4-PBI promoter (1), DNA encoding the K. lactis α-mating factor (α-MF) secretion domain (for secreted expression), a universal multiple cloning site (MCS), the K. lactis LAC4 transcription terminator (TT), and a fungal acetamidase selectable marker gene (amdS) expressed from the yeast ADH1 promoter (P_ADH1). An E. coli replication origin (ori) and ampicillin resistance gene (Ap_R) are present for propagation of pKLAC2 in E. coli.

The sequence of the pKLAC2 vector (GenBank #EU196354) and additional pKLAC2 information are available at DNA Sequences and Maps.

DNASU and Addgene are central repositories for plasmid clones and collections that may also be helpful.

Highlights
- P_LAC4-PBI promoter does not express in E. coli allowing toxic genes to be cloned prior to their expression in yeast.
- Universal MCS lies downstream of α-MF secretion domain and P_LAC4-PBI promoter.
- Acetamidase expression for non-antibiotic selection in K. lactis.
- Ampicillin resistance for propagation in E. coli. (See notes.)
Product Categories:

Discontinued Products
- Properties & Usage
  
  Affinity Tag
  Other
  Storage Temperature
  -20°C
  Sequence Files
  Fasta GenBank
- Related Products
  Companion Products
  
  Yeast Carbon Base Medium Powder
  
  K. lactis GG799 Competent Cells
  
  NEB® 5-alpha Competent E. coli (High Efficiency)
  
  NEB® 5-alpha Competent E. coli (Subcloning Efficiency)
  
  NEB® 5-alpha Electrocompetent E. coli
  
  K. lactis Protein Expression Kit
  
  BstXI
  
  SacII
- Product Notes
  NEB5-alpha Competent E.coli (High Efficiency) (NEB #C2987), NEB 5-alpha Electrocompetent E. coli (NEB #C2989) and NEB 5-alpha Competent E. coli (Subcloning Efficiency) (NEB #C2988J) are all recommended for propagation and subcloning this vector.
- References
  Colussi, P.A. and Taron, C.H. (2005). Appl. Environ. Microbiol.. 71, 7092-7098.
Protocols, Manuals & Usage
- Application Notes
  Coexpression of Multiple Proteins in Kluyveromyces lactis N3742
  
  Improved method for assembly of linear yeast expression N3742
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  N3742Datasheet-Lot0011210
  
  N3742Datasheet-Lot0011307
Tools & Resources
- Web Tools
  DNA Sequences and Maps Tool
Quality, Safety & Legal
- Quality Control Assays
  
  Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  pKLAC2 Vector
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)
  
  N3742Datasheet-Lot0011210
  
  N3742Datasheet-Lot0011307
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
  
  This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
  Licenses
  NOTICE TO BUYER/USER: The vector pKLAC2 is a component of an expression system that was developed from basic research at New England Biolabs, Inc. and DSM Biologics Company B.V. The buyer/user has a non-exclusive sublicense to use this system or any component thereof, including vector pKLAC2, for RESEARCH PURPOSES ONLY. A license to use this system for manufacture of clinical grade material or commercial purposes is available from New England Biolabs, Inc., or DSM Biologics Company B.V.