Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000765 | pLenti-CMV-MCS-GFP-SV-puro | 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 pLenti-CMV-MCS-GFP-SV-puro is a third generation lentiviral vector expressing GFP and puromycin.
- Vector Name:
- pLenti-CMV-MCS-GFP-SV-puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8455 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Promoter:
- RSV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- GAGCACCCAGTCCGCCCTG
pLenti-CMV-MCS-GFP-SV-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.
References
- Witwicka H, Hwang SY, Reyes-Gutierrez P, Jia H, Odgren PE, Donahue LR, Birnbaum MJ, Odgren PR. Studies of OC-STAMP in Osteoclast Fusion: A New Knockout Mouse Model, Rescue of Cell Fusion, and Transmembrane Topology. PLoS One. 2015 Jun 4;10(6):e0128275. doi: 10.1371/journal.pone.0128275. PMID: 26042409; PMCID: PMC4456411.
pLenti-CMV-MCS-GFP-SV-puro vector Sequence
LOCUS V000765 8455 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000765
VERSION V000765
KEYWORDS pLenti-CMV-MCS-GFP-SV-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8455)
AUTHORS Witwicka H, Hwang SY, Reyes-Gutierrez P, Jia H, Odgren PE, Donahue
LR, Birnbaum MJ, Odgren PR
TITLE Studies of OC-STAMP in Osteoclast Fusion: A New Knockout Mouse
Model, Rescue of Cell Fusion, and Transmembrane Topology.
JOURNAL PLoS One. 2015 Jun 4;10(6):e0128275. doi:
10.1371/journal.pone.0128275. eCollection 2015.
PUBMED 26042409
REFERENCE 2 (bases 1 to 8455)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8455)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
date: "2015-06-4"; pages: "
10.1371/journal.pone.0128275. eCollection 2015"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8455
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 113..134
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 149..179
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 187..203
/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 211..227
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 248..266
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 294..520
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 521..701
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 748..873
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1366..1599
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1784..1828
/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 1977..2018
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2095..2211
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2234..2537
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2538..2741
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2738..2762
/label="LNCX"
/note="Human CMV promoter, forward primer"
CDS 2818..3534
/label="EGFP"
/note="enhanced GFP"
promoter 3587..3916
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 3926..4522
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 4592..5180
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5267..5500
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5572..5706
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 5733..5868
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(5889..5907)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5917..5933)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6075..6530
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6556..6660
/label="AmpR promoter"
CDS 6661..7518
/label="AmpR"
/note="beta-lactamase"
rep_origin 7692..8280
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8434..8451
/label="L4440"
/note="L4440 vector, forward primer"