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pGIBS-TRP phagemid in E. coli

87485

The insert for this clone is a PCR amplified trp gene fragment from Bacillus subtilis including nucleotides 1883-4400 of Genbank K01391, and can be used as an expression control clone for microarrays. An artificial polyA tail was added onto the 3’ end of the insert and can be excised using BamHI + NotI. To excise the insert without the polyA tail, use XhoI + BamHI.

Organism
Bacillus subtilis subsp. subtilis (Ehrenberg) Cohn
Clone type
Clone
Applications
Molecular biology
Product format
Freeze-dried
Shipping information
Escherichia coli containing the plasmid
Storage conditions
2°C to 8°C
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
For newer versions of the microarray control probes, please see a set of clones genes: ATCC 87840 (kit).

Characteristics

Comments
For newer versions of the microarray control probes, please see a set of clones genes: ATCC 87840 (kit).
The insert is a PCR amplified B. subtilis trp gene fragment (Genbank KO1391 from nt 1883 - 4400), with an added artificial polyA tail. This insert was then cloned into the XhoI->NotI sites of pBluescript II KS+.
The insert contains the following restriction sites (approximate kb from the 5' end): BglII--1.25; PstI--2.06; XbaI--2.25.
Restriction digests of the clone give the following sizes (kb): NotI--5.6; NotI/XhoI--3.0, 2.8; XhoI--5.6.
Mycoplasma contamination
Not detected

Vector information

Construct size (kb)
5.4
Intact vector size
2.961
Vector name
pBluescriptII KS-
Type of vector
phagemid
Vector end
XhoI; NotI
Vector information
Excise insert: XhoI+NotI
Promoter giving the sense strand:T3
Promoter giving the antisense strand: T7
Markers
ampR
Promoters
T3; T7

Insert information

Insert size (kb)
2.5
Insert information
Gene: trpCDEF
Source: Bacillus subtilis
Genbank accession: K01391 (Nucleotides 1-2518 of the insert correspond to nucleotides 1883-4400 of K01391.)
Insert end (5’): XhoI linker
Insert end (3’): BamHI/polyA/NotI linker

Handling information

Host
Escherichia coli DH5
Medium
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). 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 give the following sizes (kb): NotI--5.6;
NotI/XhoI--3.0, 2.8; XhoI--5.6. Recommendation for verification: XhoI; NotI:
XhoI/NotI; PstI; XbaI.
- ATCC staff

The insert contains the following restriction sites (approximate kb from the 5'
end): BglII--1.25; PstI--2.06; XbaI--2.25.
- GenBank/EMBL/DDBJ

PCR amplified B. subtilis trp gene fragment (KO1391 from 1883->4400), added
artificial polyA tail and inserted into XhoI->NotI pBluescript II KS+.
- personal communication

History

Depositors
M Follettie
Special collection
NCRR Contract
Cross references
GenBank K01391

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.

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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

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