pOJ31

77416 ^™

This is a medium copy number shuttle vector that allows lacZα complementation. It was constructed from the cosmid vector pFD304 by addition of a lacZα polylinker fragment and a 2.4 kb PstI-SalI fragement containing the JHJ-3 replicon. The JHJ-3 replicon permits replication in the following actinomycete genera: Streptomyces, Saccharopolyspora and Amycolatopsis. The ClaI* site is methylated, making the ClaI site in the polylinker unique in E. coli dam+ strains. The order of the major features in the plasmid is: JHJ-3 ori ? neo ? ClaI* - synthetic promoter ? pMB1 ori ? cos site ? lac promoter ? SpeI/MCS/XbaI ? lacZ.- FEMS Microbiol. Lett. 78: 257-260, 1992.

Clone type: Vector
Applications: Molecular biology
Product format: Frozen
Shipping information: Escherichia coli containing the cosmid
Storage conditions: -80°C or colder

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Documentation

Product sheet

BSL 1

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.

General

Specific applications: expression vector
shuttle vector
vector permitting RNA synthesis in vitro
vector permitting visual detection of recombinants

Characteristics

Comments: Restriction digests of the clone give the following sizes (kb): BamHI--5.1; EcoRI--5.1; PstI/SalI--4.7, 0.3.
The JHJ-3 replicon permits replication in the following actinomycete genera: Streptomyces, Saccharopolyspora and Amycolatopsis.
The ClaI* site is methylated, making the ClaI site in the polylinker unique in E. coli dam+ strains.
IMPORTANT: To prevent amplification of a rearranged and/or deleted cosmid, we recommend streaking on LB + amp plates at 30C and picking small colonies for liquid culture.
Medium copy number shuttle vector allowing lacZalpha complementation.
Constructed from the cosmid vector pFD304 by addition of a lacZalpha-polylinker fragment and a 2.4 kb PstI-SalI fragment containing the JHJ-3 replicon.
The order of the major features in the plasmid is: JHJ-3 ori - neo - ClaI* - synthetic promoter - pMB1 ori - cos site - lac promoter - SpeI/MCS/XbaI - lacZ.
Mycoplasma contamination: Not detected

Vector information

Construct size (kb): 5.0
Vector name: pOJ31 (cosmid)
Construction: pFD304, pUC21, phage JHJ-3
Insert detection: lacZ'
Markers: kanR
Promoters: lac; T7
Replicon: pMB1; JHJ-3

Handling information

Medium: ATCC Medium 1236: LB Medium (ATCC medium 1065) with 25 mcg/ml kanamycin
Temperature: 30°C
Handling notes: Restriction digests of the clone gave the following sizes (in kb): BamHI 5.1 ; EcoRI 5.1 ; PstI/SalI 4.7, 0.3. ATCC Staff

History

Depositors: CV Dery

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.

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References

Curated Citations

Gaudreau LR, et al. Construction of a shuttle lacZ alpha-based Escherichia coli-actinomycetes vector containing the phage JHJ-3 replicon. FEMS Microbiol. Lett. 78: 257-260, 1992. PubMed: 1490607

Gaudreau LR, Dery CV. A cloned replicon of Saccharopolyspora phages JHJ-1 and JHJ-3 is stably maintained as a plasmid in various actinomycetes. Gene 126: 141-146, 1993. PubMed: 8472955

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