Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001783 | pLenti CMV/TO eGFP Puro (w159-1) | 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:
- pLenti CMV/TO eGFP Puro (w159-1)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8771 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- mPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMVforw
pLenti CMV/TO eGFP Puro (w159-1) vector 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.
pLenti CMV/TO eGFP Puro (w159-1) vector Sequence
LOCUS V001783 8771 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001783
VERSION V001783
KEYWORDS pLenti CMV/TO eGFP Puro (w159-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8771)
AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
Campisi J, Yaswen P, Cooper PK, Kaufman PD.
TITLE A versatile viral system for expression and depletion of proteins in
mammalian cells.
JOURNAL PLoS One. 2009 Aug 6;4(8):e6529.
PUBMED 19657394
REFERENCE 2 (bases 1 to 8771)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8771)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8771
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 27..131
/label="AmpR promoter"
CDS 132..989
/label="AmpR"
/note="beta-lactamase"
rep_origin 1163..1751
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1905..1922
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 2039..2060
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2075..2105
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2113..2129
/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 2137..2153
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 2174..2192
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2220..2447
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 2448..2628
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2672..2797
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3290..3523
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 3707..3751
/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 3900..3941
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 4012..4129
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 4152..4455
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 4456..4659
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4656..4680
/label="LNCX"
/note="Human CMV promoter, forward primer"
protein_bind 4764..4788
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
CDS 4854..5570
/label="EGFP"
/note="enhanced GFP"
protein_bind complement(5598..5622)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 5653..6241
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
promoter 6255..6754
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 6775..7371
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 7512..7745
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 7817..7951
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 7957..8092
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(8125..8143)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8153..8169)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 8314..8769
/direction=RIGHT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"