Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V009302 | PBAD1030A | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pBAD1030A plasmid is an arabinose-inducible expression vector with a wild-type RSF1030 origin. Its ampicillin-resistance cassette is in the same direction as the araBAD promoter, enabling controlled gene expression in E. coli.
- Vector Name:
- PBAD1030A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3586 bp
- Type:
- Cloning vector
- Replication origin:
- RSF ori
- Source/Author:
- Chakravartty V, Cronan JE.
- Promoter:
- araBAD
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
PBAD1030A vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- A Series of Medium and High Copy Number Arabinose-Inducible Escherichia coli Expression Vectors Compatible with pBR322 and pACYC184 Plasmid. 2015 September ; 81: 21–26. doi:10.1016/j.plasmid.2015.03.001.
PBAD1030A vector Sequence
LOCUS Exported 3586 bp DNA circular SYN 29-NOV-2023
DEFINITION Cloning vector PBAD1030A, complete sequence.
ACCESSION KP899255
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3586)
AUTHORS Chakravartty V, Cronan JE.
TITLE A series of medium and high copy number arabinose-inducible
Escherichia coli expression vectors compatible with pBR322 and
pACYC184
JOURNAL Plasmid 81, 21-26 (2015)
PUBMED 26021570
REFERENCE 2 (bases 1 to 3586)
AUTHORS Chakravartty V, Cronan JE.
TITLE Direct Submission
JOURNAL Submitted (06-MAR-2015) Microbiology, University of Illinois, 601S.
Goodwin Ave, Urbana, IL 61801, USA
REFERENCE 3 (bases 1 to 3586)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3586)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 81,
21-26 (2015)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-MAR-2015) Microbiology, University of Illinois, 601S. Goodwin
Ave, Urbana, IL 61801, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3586
/mol_type="other DNA"
/db_xref="taxon:1670622"
/organism="Cloning vector PBAD1030A"
CDS complement(96..974)
/codon_start=1
/gene="araC"
/product="AraC"
/label=araC
/note="transcription factor"
/protein_id="AKM76960.1"
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
gene complement(96..974)
/gene="araC"
/label=araC
promoter 1001..1285
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
terminator 1569..1655
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1747..1774
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
misc_feature complement(1834..2462)
/label=RSF ori
/note="Plasmids containing the RSF 1030 origin
ofreplication can be propagated in E. coli cells that
contain additional plasmids with compatible origins."
promoter 2479..2583
/gene="bla"
/label=AmpR promoter
CDS 2584..3444
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/note="ampicillin resistance"
/protein_id="AKM76961.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 2584..3444
/gene="bla"
/label=bla