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K. lactis Vectors >
pKLAC1 Vector |
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- PLAC4-PBI promoter does not express in E. coli allowing toxic genes to be cloned prior to their expression in yeast.
- Convenient MCS lies downstream of α-MF secretion domain and PLAC4-PBI promoter.
- Acetamidase expression for non-antibiotic selection in K. lactis.
- Ampicillin resistance for propagation in E. coli.
Description:
The vector pKLAC1 directs high-level expression of a recombinant protein from the yeast Kluyveromyces lactis and is part of the K. lactis Protein Expression Kit (NEB #E1000). pKLAC1 can be used for either intracellular or secreted protein expression. SacII or BstXI linearized pKLAC1 integrates into the LAC4 locus of the K. lactis genome upon transformation of K. lactis GG799 Competent Cells (NEB #C1001).
Vector pKLAC1 contains the strong K. lactis PLAC4-PBI promoter (1), DNA encoding the K. lactis α-mating factor (α-MF) secretion domain (for secreted expression), a multiple cloning site (MCS), the K. lactis LAC4 transcription terminator (TT), and a fungal acetamidase selectable marker gene (amdS) expressed from the yeast ADH2 promoter (PADH2). An E. coli replication origin (ori) and ampicillin resistance gene (ApR) are present for propagation of pKLAC1 in E. coli.
Source:
pKLAC1 is isolated from E. coli strain ER2268 by a standard DNA purification procedure.
Concentration:
1,000 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Storage Temperature:
-20°C
pKLAC1 plasmid map. Unique restriction sites are shown in bold. SacII and BstXI sites are shown in red.
pKLAC1 multiple cloning site (MCS). The K. lactis α-mating factor secretion domain is shown with a blue background. Only unique restriction sites are shown.
References
- Colussi, P.A. and Taron, C.H., unpublished observations.
Companion Products
α-Mating Factor Sequencing Primer (18-mer)
K. lactis GG799 Competent Cells
K. lactis Protein Expression Kit
BstXI
Integration Primer Set
LAC4mt Promoter Sequencing Primer (18-mer)
pKLAC1 Reverse Sequencing Primer (18-mer)
SacII
Yeast Medium Pack
Legal
Licenses/Patents/Disclaimers:
The vector pKLAC1 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 pKLAC1, 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.
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