Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011544 | pGL3-Basic | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pGL3-Basic is a promoterless vector that can be used to measure the activity of promoter and enhancer sequences with a luciferase assay.
- Vector Name:
- pGL3-Basic
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4818 bp
- Type:
- Luciferase Vectors
- Replication origin:
- ori
- Source/Author:
- Promega
- Copy Number:
- High copy number
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pGL3-Basic 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
- Madlala P, Mkhize Z, Naicker S, Khathi SP, Maikoo S, Gopee K, Dong KL, Ndung'u T. Genetic variation of the HIV-1 subtype C transmitted/founder viruses long terminal repeat elements and the impact on transcription activation potential and clinical disease outcomes. PLoS Pathog. 2023 Jun 12;19(6):e1011194. doi: 10.1371/journal.ppat.1011194. PMID: 37307292; PMCID: PMC10289673.
pGL3-Basic vector Sequence
LOCUS 40924_21953 4818 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pGL3-Basic, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4818)
AUTHORS Groskreutz DJ, Schenborn ET.
TITLE Direct Submission
JOURNAL Submitted (26-JAN-1996) D.J. Groskreutz, R
REFERENCE 2 (bases 1 to 4818)
AUTHORS Kenefick K.
TITLE Direct Submission
JOURNAL Submitted (05-MAR-2001) Technical Writing, Promega Corporation, 2800
Woods Hollow Road, Madison, WI 53711-5399, USA
REFERENCE 3 (bases 1 to 4818)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4818)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(26-JAN-1996) D.J. Groskreutz, R"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(05-MAR-2001) Technical Writing, Promega Corporation, 2800 Woods
Hollow Road, Madison, WI 53711-5399, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT On Mar 5, 2001 this sequence version replaced U47295.1.
FEATURES Location/Qualifiers
source 1..4818
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..58
/label=multiple cloning site
/note="multiple cloning site"
CDS 88..1737
/codon_start=1
/label=luciferase
/note="firefly luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
polyA_signal complement(1781..1902)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2061..2080)
/label=RV primer4 sequencing primer binding site
/note="RV primer4 sequencing primer binding site"
rep_origin 2318
/label=ColE1-derived plasmid replication origin
/note="ColE1-derived plasmid replication origin"
rep_origin complement(2321..2909)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3083..3940)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3941..4045)
/label=AmpR promoter
rep_origin 4072..4527
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal 4658..4706
/label=poly(A) signal
/note="synthetic polyadenylation signal"
misc_feature 4720..4811
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"