Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V014385 | pLV-H1-EF1a-Puro | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pLV-H1-EF1a-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8415 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Temperature:
- 37℃
pLV-H1-EF1a-Puro 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.
pLV-H1-EF1a-Puro vector Sequence
LOCUS V014385 8415 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014385
VERSION V014385
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8415)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8415
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 9..235
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 463..588
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1081..1314
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1499..1543
/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 1692..1733
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter complement(1882..2096)
/label="H1 promoter"
/note="human H1 RNA promoter"
misc_feature 2116..2232
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2437..3618
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3653..4249
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 4267..4855
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4942..5175
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5247..5381
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 5408..5543
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(5564..5582)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5592..5608)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5750..6205
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6231..6335
/label="AmpR promoter"
CDS 6336..7193
/label="AmpR"
/note="beta-lactamase"
rep_origin 7367..7955
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 8243..8264
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8279..8309
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 8317..8333
/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)."
primer_bind 8341..8357
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 8378..8396
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"