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pDMS197

87694

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
Counterselection levansucrase
Suicide vector
Vector permitting allelic exchange of cloned inserts

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): XbaI-- 6.55; KpnI-- 6.55; HindIII-- 2.9, 2.2, 1.4.

The sacB1 allele is a variant of sacB with certain restriction sites removed by site directed mutagenesis. Expression of sacB confers sensitivity to sucrose.

The conditional R6K origin of replication requires that the pi protein be expressed in trans for plasmid maintenance.

One of four allelic exchange suicide vectors (ATCC 87691 - 87694) that provide both selection for chromosomal integration (ampR, cmlR, kanR or tetR) and counterselection for loss of vector DNA and the wild type allele.

Construct is suicide plasmid in any host not expressing pir.

Cloned inserts may be integrated into the host chromosome (a single recombination event) following electroporation and appropriate antibiotic selection.

Negative selection for sucrose sensitivity (sacB) selects for a second recombination event resulting in loss of vector DNA.
Mycoplasma contamination
Not detected

Vector information

Vector name
pDMS197 (plasmid)
Type of vector
plasmid
Construction
pRE107, tetR (pBR322)
Host range
broad host range
Cloning sites
BamHI; XbaI; KpnI; SacI; SmaI
Markers
sacB; sacB1; tetR
MCS
BamHI...EcoRI, ->
Polylinker sites
BamHI; HindIII; XbaI; KpnI; SphI; EcoRV; SacI; SmaI; EcoRI
Replicon
conditional R6K ori; oriT
Restriction sites
EcoRI; HindIII

Insert information

Target gene
levansucrase

Handling information

Host
Escherichia coli SM10 lambda pir
Medium
Temperature
37°C
Handling notes
Restriction digests of the clone give the following sizes (kb): XbaI-- 6.55;
KpnI-- 6.55; HindIII-- 2.9, 2.2, 1.4.
- ATCC staff

One of four allelic exchange suicide vectors (ATCC 87691 - 87694) that provide
both selection for chromosomal integration (ampR, cmLR, kanR or tetR) and
counterselection for loss of vector DNA and the wild type allele.
- Gene 207: 149-157, 1998

Construct is suicide plasmid in any host not expressing pir.
- Gene 207: 149-157, 1998

The conditional R6K origin of replication requires that the pi protein be
expressed in trans for plasmid maintenance.
- Cell 15: 1199-1208, 1978

Cloned inserts may be integrated into the host chromosome (a single
recombination event) following electroporation and appropriate antibiotic
selection.
- Gene 207: 149-157, 1998

Negative selection for sucrose sensitivity (sacB) selects for a second
recombination event resulting in loss of vector DNA.
- Infect. Immun. 59: 4310-4317, 1991
- Mol. Microbiol. 5: 1447-1457, 1991

The sacB1 allele is a variant of sacB with certain restriction sites removed by
site directed mutagenesis. Expression of sacB confers sensitivity to sucrose.
- Mol. Microbiol. 18: 877-889, 1995

History

Depositors
DM Schifferli
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

Edwards RA, et al. Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Gene 207: 149-157, 1998. PubMed: 9511756

Donnenberg MS, Kaper JB. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect. Immun. 59: 4310-4317, 1991. PubMed: 1937792

Blomfield IC, et al. Allelic exchange in Escherichia coli using the Bacillus subtilis sacB gene and a temperature-sensitive pSC101 replicon. Mol. Microbiol. 5: 1447-1457, 1991. PubMed: 1686293

Kolter R, et al. Trans-complementation-dependent replication of a low molecular weight origin fragment from plasmid R6K. Cell 15: 1199-1208, 1978. PubMed: 728998

McIver KS, et al. The elastase propeptide functions as an intramolecular chaperone required for elastase activity and secretion in Pseudomonas aeruginosa. Mol. Microbiol. 18: 877-889, 1995. PubMed: 8825092

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