Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000707 | 1.3xWT HBV-Luciferase | 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
The 1.3xWT HBV - Luciferase vector is an important tool for studying hepatitis B virus (HBV) gene expression.
It is constructed by replacing specific sequences in the HBV genome with the luciferase gene, enabling the measurement of HBV transcriptional activity through luciferase activity. Its key characteristics include a specific start codon from the Core ORF and regulation by the pgRNA promoter, ensuring accurate reflection of HBV gene expression changes. The main advantages are its high specificity and sensitivity in detecting HBV transcriptional activity and its usefulness in studying the roles of different regulatory elements in the HBV genome.
It should be chosen when investigating HBV gene expression regulation, particularly when analyzing the effects of various factors on HBV transcription or when exploring the interactions between different regulatory elements within the HBV genome.
- Vector Name:
- 1.3xWT HBV-Luciferase
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6656 bp
- Type:
- Mammalian Expression, Luciferase
- Replication origin:
- ori
- Promoter:
- SP6
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
1.3xWT HBV-Luciferase 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
- Doitsh G, Shaul Y. Enhancer I predominance in hepatitis B virus gene expression. Mol Cell Biol. 2004 Feb;24(4):1799-808. doi: 10.1128/MCB.24.4.1799-1808.2004. PMID: 14749394; PMCID: PMC344184.
1.3xWT HBV-Luciferase vector Sequence
LOCUS V000707 6656 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000707
VERSION V000707
KEYWORDS 1.3xWT HBV-Luciferase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6656)
AUTHORS Doitsh G, Shaul Y
TITLE Enhancer I predominance in hepatitis B virus gene expression.
JOURNAL Mol Cell Biol. 2004 Feb;24(4):1799-808.
PUBMED 14749394
REFERENCE 2 (bases 1 to 6656)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6656)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell
Biol."; date: "2004-02"; volume: "24(4)"; pages: "1799-808"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6656
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 355..816
/gene="X"
/label="Protein X"
/note="Protein X from Hepatitis B virus genotype A2 subtype
adw2 (strain Rutter 1979). Accession#: P69713"
CDS 969..2618
/label="luciferase"
/note="firefly luciferase"
CDS 3317..3778
/gene="X"
/label="Protein X"
/note="Protein X from Hepatitis B virus genotype A2 subtype
adw2 (strain Rutter 1979). Accession#: P69713"
promoter complement(3981..3999)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4017..4033)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4041..4057)
/label="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4065..4095)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(4110..4131)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4248..4265)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(4419..5007)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5181..6038)
/label="AmpR"
/note="beta-lactamase"
promoter complement(6039..6143)
/label="AmpR promoter"
primer_bind 6211..6229
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6267..6289)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind 6389..6408
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6617..6633
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 6640..6656
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"