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pKD136, pKD8.243A plasmids in E. coli

98014

There are 2 plasmids in this strain, pKD136 and pKD8.243A. They are ampicillin and chloramphenicol resistant respectively.This strain supports a portion of the biocatalytic conversion of D-glucose into adipate (a reagent in the production of nylon).  It converts D-glucose into catechol.  Colonies turn brown as catechol accumulates.  The transketolase (tkt) coding sequence can be recovered as a 2.4 kb EcoRI fragment.  The aroF coding sequence can be recovered as a 5.0 kb BamHI fragment.  The aroB coding sequence can be recovered as a 1.65 kb SphI fragment.
Organism
Klebsiella pneumoniae (Schroeter) Trevisan
Clone type
Clone
Applications
Molecular biology
Product format
Freeze-dried
Shipping information
Escherichia coli containing the plasmid
Storage conditions
2°C to 8°C
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Documentation

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Biosafety Icon 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.

Detailed product information

General

Specific applications
produces protein 3-dehydroquinate synthase
produces protein 3-dehydroshikimate dehydratase
produces protein 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase
produces protein protocatechuate decarboxylase
produces protein transketolase

Characteristics

Comments
Restriction digests of the plasmids in this strain give the following sizes (kb): BamHI--10.0, 5.0, 4.8, 3.4, 1.1, 0.3; EcoRI--9.2, 8.8, 4.0, 2.4; SphI--18.0, 4.4, 1.8.
This strain supports a portion of the biocatalytic conversion of D-glucose into adipate (a reagent in the production of nylon). It converts D-glucose into catechol. Colonies turn brown as catechol accumulates.
The tkt CDS can be recovered as a 2.4 kb EcoRI fragment. The aroF CDS can be recovered as a 5.0 kb BamHI fragment.
The aroB CDS can be recovered as a 1.65 kb SphI fragment.
Mycoplasma contamination
Not detected
Technical information
ATCC Technical Services does not have technical information on patent deposits that are not produced or characterized by ATCC. Additional information can be found in the corresponding patent available from the patent holder or with the U.S. and/or international patent office.

Vector information

Vector name
pKD8.243A: unknown // pKD136: pBR325
Vector information
Information on plasmid #1: pKD8.243A
Size (kb): Unknown
Marker: cmlR

Insert component:
Insert size (kb):

Handling information

Host
Escherichia coli AB2834
Temperature
37°C
Handling procedure
  1. Open vial according to instructions.
  2. Asceptically add 0.3 to 0.4 mL of liquid medium to the freeze-dried pellet and mix well.  Transfer 100 µL to a test tube containing 5 mL LB+ ampicillin (50-100 µg/mL) and chloramphenicol (50 ug/mL). A loopful of culture can also be streaked on an agar plate of the same. Incubate cultures at 37°C.
  3. Isolate DNA using standard plasmid preparation procedures.
Handling notes
Restriction digests of the clone gave the following sizes   (in kb):  BamHI - 10.0, 5.0, 4.8, 3.4, 1.1, 0.3 ; EcoRI - 9.2, 8.8, 4.0, 2.4  ; SphI -  18.0, 4.4, 1.8.      -ATCC Staff

History

Depositors
Purdue Univ. Res. Fndn., A Berry, Purdue Univ. Res. Fndn.
Patent depository
This material was deposited with the ATCC Patent Depository to fulfill U.S. or international patent requirements. This material may not have been produced or characterized by ATCC.  As an International Depository Authority (IDA) for patent deposits, ATCC is required to complete viability testing only at time of initial deposit of patent material. Patent deposits are made available on behalf of the Depositor when the pertinent U.S. or international patent is issued, but material may not be used to infringe the patent claims.

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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.
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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.

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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.

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Disclosures
This material is cited in a US and/or international patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Depositor of the party to which the material was furnished.

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

Draths KM, Frost JW. Genomic direction of synthesis during plasmid-based biocatalysis. J. Am. Chem. Soc. 112: 9630-9632, 1990.

Draths KM, Frost JW. Synthesis using plasmid-based biocatalysis: plasmid assembly and 3-deoxy-D-arabino-heptulosonate production. J. Am. Chem. Soc. 112: 1657-1659, 1990.

Draths KM, Frost JW. Environmentally compatible synthesis of adipic acid from D-glucose. J. Am. Chem. Soc. 116: 399-400, 1994.

Frost JW. Enhanced production of common aromatic pathway compounds. US Patent 5,168,056 dated Dec 1 1992

Pittard J, Wallace BJ. Distribution and function of genes concerned with aromatic biosynthesis in Escherichia coli. J. Bacteriol. 91: 1494-1508, 1966. PubMed: 5326114

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