pBAD30

87401 ^™

This vector is one of several tightly controlled expression vectors (ATCC#87393-87402) regulated by the arabinose operon. Vectors differ in replicon, antibiotic resistance marker, multiple cloning site and need for translation initiation sequences. Cultures should be grown in minimal media for more reproducible induction of expression. Expression is induced in glycerol-containing media by the addition of arabinose. Expression is repressed by addition of glucose or other catabolites. Plasmid copy number is low due to the p15A replicon. This vector can be used when reduced gene expression is desirable. Plasmid is compatible with pBR-derived plasmids and may be used for coexpression of cloned inserts. Plasmid contains the 3' portion of the cat (cmLR) gene which could promote recombination if this plasmid is used in combination with other compatible cmLR plasmids. Recombination can be avoided by the use of recA host strains, or it can be used to advantage to intentionally exchange markers among plasmids. The following primers can be used for sequencing of cloned inserts: 5' primer (27 ? 8 bp upstream of the NheI site):5? ? CTGTTTCTCCATACCCGTT- 3'; and one of two 3' primers: 3' primer 1 (2-19 bp downstream of the HindIII site): 5' - CTCATCCGCCAAAACAG-3'; 3' primer 2 (17-33 bp downstream of the HindIII site):5' -GGCTGAAAATCTTCTCT-3'.

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

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Documentation

Product sheet

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.

General

Specific applications: expression vector
produces protein arabinose regulator
vector containing primer sites useful for sequencing
vector with low copy number

Characteristics

Comments: Restriction digests of the clone give the following sizes (kb): AvaI--2.0, 1.85, 1.2; BamHI--5.0; PstI--4.0, 1.1.
Cultures should be grown in minimal media for more reproducible induction of expression. Expression is induced in glycerol-containing media by addition of arabinose. Expression is repressed by addition of glucose or other catabolites.
One of several tightly controlled expression vectors (ATCC 87393-87402) regulated by the arabinose operon. The vectors differ in replicon, antibiotic resistance marker, multiple cloning site and mechanism of initiation of translation.
The following primers can be used for sequencing of cloned inserts: 5' primer (27 - 8 bp upstream of the NheI site) 5'-CTGTTTCTCCATACCCGTT-3'; and one of two 3' primers: 3' primer 1 (2 - 19 bp downstream of the HindIII site) 5'-CTCATCCGCCAAAACAG-3';
3' primer 2 (17 - 33 bp downstream of the HindIII site) 5'-GGCTGAAAATCTTCTCT-3'.
Plasmid copy number is low due to the p15A replicon. This vector can be used when reduced gene expression is desirable.
Plasmid contains the 3' portion of the cat (cmlR) gene which could promoter recombination if this plasmid is used in combination with other compatible cmlR plasmids.
Plasmid is compatible with pBR-derived plasmids and may be used for coexpression of cloned inserts.
Recombination can be avoided by the use of recA host strains, or it can be used to advantage to intentionally exchange markers among plasmids.
Mycoplasma contamination: Not detected

Vector information

Construct size (kb): 4.923
Vector name: pBAD30 (phagemid)
Type of vector: phagemid
Construction: pBAD28
Vector information: other: CAP site
Markers: araC; ampR
MCS: NheI...HindIII
Operator: I2 + I1; O1; O2
Promoters: araC; arabinose BAD
Regulator: araC
Replicon: M13; p15A
Terminator: rrnB T1 + T2
Transcription terminator: rrnB T1; rrnB T2

Insert information

Target gene: arabinose regulator

Handling information

Medium: ATCC Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Temperature: 37°C
Handling notes: Restriction digests of the vector gave the following sizes (in kb): BamHI 5.0 ; PstI 4.0, 1.1; AvaI 2.0, 1.85. ATCC Staff

History

Depositors: L Guzman, J Beckwith
Special collection: NCRR Contract

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.

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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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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

Guzman LM, et al. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J. Bacteriol. 177: 4121-4130, 1995. PubMed: 7608087

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