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pNK2883

77339

Organism
Escherichia coli (Migula) Castellani and Chalmers
Classification
Enterobacteriaceae, Escherichia
Clone type
Clone
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
produces protein transposase, Tn10, ats1 ats2 (ATS)

Characteristics

Comments
This cassette is inserted into the HindIII site (original nt 2272) of the ATS transposase-containing EcoRI fragment of pNK2881, ATCC 77351.
The mini-Tn10 tetR cassette is unstable because a portion of the tetR marker inside the transposon is directly repeated in adjacent sequences outside the transposon.
The plasmid should be propagaged in a recA- host, and the presence of the 2.9 kb BamHI fragment containing the cassette should be verified before use.
The tetR marker SHOULD NOT be used for selection, as it is not effective in multiple copies.
Construct useful for general transposon mutagenesis in Escherichia coli.
pBR322 was modified by deletion of bp 75-2352 and destruction (by filling in) of the HindIII site.
Contains the ats1 ats2 transposase gene that permits relaxed insertion specificity (altered target specificity, ATS). Expression is regulated by the Ptac promoter, inducible by IPTG.
The transposase segment extends from IS10R to the EcoRI site at nt 3140 of Tn10, with a deletion of nucleotides 1329-1942 to remove the transposase binding site.
Contains a 2.9 kb mini-Tn10 cassette conferring tetracycline resistance, bounded by inverted repeats of the outermost 70 bp of IS10R and embedded in 40 bp of lambdacI terminating in HindIII sites.
Transcription from the Ptac promoter is in a direction opposite to that of amp.
The order of the major features in this plasmid is: EcoRI - Ptac - ATS transposase - mini-Tn10 tetA tetR - EcoRI - pMB1 ori - ampR.
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
7.599999904632568

Insert information

Insert size (kb)
5.5
Type of DNA
genomic
Target gene
transposase; Tn10; ats1 ats2 (ATS)
Gene product
transposase, Tn10, ats1 ats2 (ATS)

Handling information

History

Depositors
N Kleckner

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

Kleckner N, et al. Uses of transposons with emphasis on Tn10. Methods Enzymol. 204: 139-180, 1991. PubMed: 1658561

Nancy Kleckner, personal communication

For product-related inquiries and issues, contact Product Experience:

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