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p34E-Tp plasmid in E. coli

87447

This plasmid contains a trimethoprim resistance cassette which is useful as a selectable marker, for cassette mutagenesis of cloned genes, and for the construction of new cloning vectors. The cassette insert was amplified using primers that allowed incorporation of two EcoRI sites, resulting in 3 bp mismatches with respect to the portion of the GenBank record J01774 described below.     This vector was constructed by inserting a trimethoprim resistance cassette into the pGEM-3/4 vector. The insert and vector components are described below under ?insert component? and ?vector component?, respectively.
Organism
R388
Clone type
Vector
Applications
Molecular biology
Product format
Frozen
Shipping information
Escherichia coli containing the plasmid in glycerol stock
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
contains easily purifiable cassette(s) for construction dihydrofolate reductase, trimethoprim resistance
produces protein dihydrofolate reductase, trimethoprim resistance

Characteristics

Comments
Restriction digests of the clone give the following sizes (kb): AvaI--2.9, 0.4, 0.2; BglI--3.5; EcoRI--2.9, 0.6.
Plasmid containing a trimethoprim resistance cassette useful as a selectable marker, for cassette mutagenesis of cloned genes, and for the construction of new cloning vectors.
Trimethoprim insert contains nucleotides 399-1016 of GenBank J01774 and was cloned into the EcoRI site of the pGEM-3/4 vector.
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
3.527
Vector name
p34E-Tp (plasmid)
Construction
pGEM-3, pGEM-4, dhfrIIb (tmpR)
Markers
tmpR; ampR
MCS
EcoRI...HindIII; HindIII...EcoRI
Promoters
Expression: lac; In vitro

Insert information

Insert size (kb)
0.61799999999999999
Type of DNA
cDNA
Target gene
dihydrofolate reductase, trimethoprim resistance
Gene product
dihydrofolate reductase, trimethoprim resistance [dhfrIIb]

Handling information

Medium
Temperature
37°C
Handling notes
Restriction digests of the clone gave the following sizes   (in kb):  AvaI – 2.9, 0.4, 0.2 ; BglI – 3.5 ; EcoRI – 2.9, 0.6.                                                                 –ATCC Staff

History

Depositors
DE Woods, D DeShazer
Special collection
NCRR Contract
Cross references
GenBank J01774

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

DeShazer D, Woods DE. Broad-host-range cloning and cassette vectors based on the R388 trimethoprim resistance gene. BioTechniques 20: 762-764, 1996. PubMed: 8723912

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