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pFD666

77286

A cosmid shuttle vector for actinomycetes or E. coli that can be used for production of ssDNA and has promoters for in vitro RNA synthesis.  The order of the major features in this plasmid is: rrnC terminator - SP6 -SphI/MCS/HindIII - T7 - trpA terminator - neo - synthetic promoter - pMB1 ori -cos - M13 ori - pJV1 ori.Terminators flanking the polylinker protect the vector from transcripts originating in cloned inserts.  - Gene 111: 115-118, 1992The pJV1 ori from Streptomyces phaeochromogenes is compatible with plasmids derived from pIJ101.  This vector can be used for complementation studies using multiple cloned genes in one host.  - Gene 111: 115-118, 1992M13K07 helper phage improves the yield of ssDNA in E. coli with this vector.  M13mp18 may also be used.  - Gene 111: 115-118, 1992
Clone type
Vector
Applications
Molecular biology
Product format
Frozen
Shipping information
Escherichia coli containing the plasmid
Storage conditions
-80°C or colder
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
shuttle vector
vector for constructing genomic libraries
vector permitting production of single-stranded DNA

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): BamHI--5.4; PstI--5.4; PvuII--3.7, 1.7; SacI--4.5, 0.9.
M13K07 helper phage improves the yield of ssDNA in E. coli with this vector. M13mp18 may also be used.
The neo promoter was replaced by a synthetic promoter recognized by both hosts.
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.
Cosmid shuttle vector permitting production of ssDNA and containing promoters for in vitro RNA synthesis. Terminators flanking the polylinker protect the vector from transcripts originating in cloned inserts.
The ori of pJV1 from Streptomyces phaeochromogenes is compatible with plasmids derived from pIJ101. This vector can be used for complementation studies using multiple cloned genes in one host.
The order of the major features in this plasmid is: rrnC terminator - SP6 - SphI/MCS/HindIII - T7 - trpA terminator - neo - synthetic promoter - pMB1 ori - cos - M13 ori - pJV1 ori.
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
5.25
Vector name
pFD666 (cosmid)
Construction
pIBI76, Lorist6, pUC118, pCOS2EMBL, pWOR126
Markers
kanR
Promoters
SP6; T7
Replicon
pMB1; M13; pJV1
Terminator
rrnC, trpA

Handling information

Medium
Temperature
30°C
Handling notes
Restriction digests of the vector gave the following sizes (kb): BamHI--5.4;PstI--5.4;PvuII--3.7, 1.7; SacI--4.5, 0.9.

History

Depositors
R Brzezinski

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

Denis F, Brzezinski R. A versatile shuttle cosmid vector for use in Escherichia coli and actinomycetes. Gene 111: 115-118, 1992. PubMed: 1547947

Denis F, Brzezinski R. An improved aminoglycoside resistance gene cassette for use in gram-negative bacteria and Streptomyces. FEMS Microbiol. Lett. 65: 261-264, 1991. PubMed: 1655560

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