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pAR1219

MBA-387

This product is an ATCC manufactured and accessioned progeny of 39563 cited in US Patent Number 4,952,496.
Organism
Escherichia coli (Migula) Castellani and Chalmers
Classification
Enterobacteriaceae, Escherichia
Clone type
Clone
Applications
Molecular biology
Product format
Frozen
Storage conditions
-80°C or colder
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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 RNA polymerase

Characteristics

Genotype
pAR1219 T7gene1 lacI F- hsdR19 recA1 rpoB331 IN(rrnD-rrnE)1 lambda-
Comments

Restriction digests of the clone give the following sizes (kb): BamHI--4.4 (doublet); BglII-uncut; EcoRI--8.8; PstI--8.8; HindIII--8.8.

The T7 RNA polymerase expression construct, encoding gene 1 and lac regulatory sequences, can easily be excised as a 4.4 kb BamHI fragment for insertion into another construct.

The insert size given is for that of the sum of the lac and gene 1 sequences.

Gene1 was sequenced from T7 wild-type DNA. The junctions between gene 1 and lac were sequenced. Lac sequence is taken from GenBank J01637.

pAR1219 produces large amounts of T7 RNA polymerase in a suitable host upon induction with IPTG. Large amounts of active T7 RNA polymerase can be purified from the strain BL21/pAR1219.

The lac fragment contains the lacI promoter, lacI gene, lacUV5 promoter, lac operator, and the translation start site and first 147 amino acids of B-galactosidase.

pAR1219 was made by inserting a 1724 bp HincII fragment from pMC1, containing lac control elements, into the BglII site of pAR1173.

The construction consists of a fragment containing the T7 RNA polymerase gene (gene 1, including nucleotides 3145 to 5841 of T7 DNA) under the transcriptional regulation of lacI and the lacUV5 promoter.

Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
8.8
Intact vector size
4.363
Vector name
pBR322
Type of vector
plasmid
Construction
pBR313
Host range
Escherichia coli
Vector end
BamHI
Vector information
Cross references: DNA Seq. Acc.: J01749
Cloning sites
EcoRI; ClaI; HindIII; EcoRV; BamHI; SphI; SalI; XmaIII; NruI; BspMI; BsmI; StyI; AvaI; BalI; BspMII; PvuII; Tth111I; NdeI; AflIII; PpaI; PstI; PvuI; ScaI; SspI; AatII
Markers
ampR; tetR
Replicon
pMB1

Insert information

Insert size (kb)
4.4400000000000004
Type of DNA
genomic
Insert information
Genomic copy number: unique
Cross references: DNA Seq. Acc.: J02518
Genome
bacteriophage T7
Target gene
RNA polymerase
Gene name
RNA polymerase
Gene product
RNA polymerase [gene 1]
Gene symbol
gene 1
Contains complete coding sequence
Unknown
Insert end
Modification: BamHI linkers

Handling information

Host
Escherichia coli HMS174; Richardson HMS 174
Medium
Temperature
37°C
Handling notes
Restriction digests of the clone give the following sizes (kb): BamHI--4.4 (doublet); BglII-uncut; EcoRI--8.8; PstI--8.8; HindIII--8.8.
- ATCC staff

The insert size given is for that of the sum of the lac and gene 1 sequences. Gene1 was sequenced from T7 wild-type DNA. The junctions between gene 1 and lac were sequenced. Lac sequence is taken from GenBank J01637. The lac fragment contains the lacI promoter, lacI gene, lacUV5 promoter, lac operator, and the translation start site and first 147 amino acids of B-galactosidase. pAR1219 produces large amounts of T7 RNA polymerase in a suitable host upon induction with IPTG. Large amounts of active T7 RNA polymerase can be purified from the
strain BL21/pAR1219. pAR1219 was made by inserting a 1724 bp HincII fragment from pMC1, containing lac control elements, into the BglII site of pAR1173. The construction consists of a fragment containing the T7 RNA polymerase gene (gene 1, including nucleotides 3145 to 5841 of T7 DNA) under the transcriptional regulation of lacI and the lacUV5 promoter. The T7 RNA polymerase expression construct, encoding gene 1 and lac regulatory sequences, can easily be excised as a 4.4 kb BamHI fragment for insertion into another construct.
- U.S. Pat. 4,952,496 dated Aug. 28, 1990

History

Deposited as
ATCC accessioned progeny of pAR1219 cited in US Patent Number 4,952,496 as 39563.
Cross references
GenBank J02518

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

Studier FW, et al. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. US Patent 4,952,496 dated Aug 28 1990

Studier FW, Moffatt BA. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J. Mol. Biol. 189: 113-130, 1986. PubMed: 3537305

Davanloo P, et al. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc. Natl. Acad. Sci. USA 81: 2035-2039, 1984. PubMed: 6371808

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