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pGK::nucMCS plasmid in Lactococcus lactis subsp. lactis

87139

Organism
Staphylococcus aureus Rosenbach
Clone type
Vector
Applications
Molecular biology
Product format
Frozen
Mission Collection Item
This is a Mission Collection Item.

Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Detailed product information

General

Specific applications
vector permitting visual detection of recombinants

Characteristics

Comments
To detect nuclease activity, use TBD agar: 100 ml 0.05 M Tris, pH 9; 1 g agar; 1 g NaCl; 10 ul CaCl2, 1 M; ssDNA (0.5% w/v). Heat until DNA and agar are dissolved and melted. Add 9 mg Toluidine blue O dye (Sigma T3260) and heat again. Melt for use.
Replicates as a rollling circle to a level of 20-30 copies per chromosome in L. lactis.
High copy-number plasmid vector for gram positive bacteria. Permits identification of recombinants by detecting secreted nuclease activity.
Phenotype(s) verified: eryR
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
4.30
Vector name
pGK::nucMCS (plasmid)
Type of vector
plasmid
Construction
pGK12, nuc
Vector information
leader peptide: nuc leader
Coding sequence
nuc (processed)
Markers
eryR
MCS
NsiI...XhoI
Promoters
nuc
Replicon
pWV01
Ribosome binding site
nuc

Insert information

Insert size (kb)
0.85799999999999998
Type of DNA
genomic
Insert information
Source of nuc gene: Staphylococcus aureus
Gene product
nuclease(micrococcal nuclease; staphylococcal nuclease) [nuc]

History

Depositors
Y Le Loir, P Langella
Special collection
NCRR Contract

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Le Loir Y, et al. Direct screening of recombinants in gram-positive bacteria using the secreted staphylococcal nuclease as a reporter. J. Bacteriol. 176: 5135-5139, 1994. PubMed: 8051029

Lachica RV, et al. Metachromatic agar diffusion methods for detecting staphyloccal nuclease activity. Appl. Microbiol. 21: 585-587, 1971. PubMed: 4930272

Kok J, et al. Construction of plasmid cloning vectors for lactis streptococci which also replicate in Bacillus subtilis and Escherichia coli. Appl. Environ. Microbiol. 48: 726-731, 1984. PubMed: 6095756

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