Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001141 | pHIV-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
pHIV-LUC is a specialized plasmid designed for molecular biology research applications. It features a unique construction where sequences from the HIV enhancer (-2453 to +80) are linked to the firefly luciferase gene. This design enables the plasmid to be used in a variety of assays to study gene expression and transcriptional regulation. When transfected into cells, it allows for the quantification of luciferase activity, which serves as an indicator of transcriptional activity.
It is a valuable tool in investigations related to DNA repair mechanisms and the effects of various factors on gene expression. In different experimental setups, changes in luciferase activity can provide insights into the functionality of the transfected cells and the impact of different treatments or conditions on the transcriptional machinery.
- Vector Name:
- pHIV-Luciferase
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8713 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- EF-1α
- 5' Primer:
- 5'-TGGAATTTGCCCTTTTTGAG-3'
- 3' Primer:
- 5'-AGGAACTGCTTCCTTCACGA-3'
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pHIV-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
- Madrid O, Varea S, Sanchez-Perez I, Gomez-Garcia L, De Miguel E, Gomez De Segura IA, Perona R. Growth hormone protects against radiotherapy-induced cell death. Eur J Endocrinol. 2002 Oct;147(4):535-41. doi: 10.1530/eje.0.1470535. PMID: 12370117.
pHIV-Luciferase vector Sequence
LOCUS V001141 8713 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001141
VERSION V001141
KEYWORDS pHIV-Luciferase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8713)
TITLE Lentivirus-EF1alpha-MCS-IRES-Luciferase
REFERENCE 2 (bases 1 to 8713)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8713)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8713
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(47..66)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 618..820
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 835..1015
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1686..1919
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2104..2148
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 2297..2338
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2446..2563
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2627..3805
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature 3866..4449
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4449..6098
/label="luciferase"
/note="firefly luciferase"
primer_bind complement(6156..6175)
/label="EBV-rev"
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6193..6209)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
LTR 6719..6899
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6961..7549)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7723..8580)
/label="AmpR"
/note="beta-lactamase"
promoter complement(8581..8685)
/label="AmpR promoter"