Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V016743 | LentiMPH v2 | 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
lentiMPH v2 is a lenti vector encoding the MS2-P65-HSF1 activator helper complex with a 2A Hygro resistance marker (EF1a-MS2-p65-HSF1-2A-Hygro-WPRE). This version has a higher virus titer than the first one.
- Vector Name:
- LentiMPH v2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11739 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Hyg,Zeo
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
LentiMPH v2 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
- Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F. Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Nat Protoc. 2017 Apr;12(4):828-863.
LentiMPH v2 vector Sequence
LOCUS V016743 11739 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016743
VERSION V016743
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11739)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11739
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(488..613)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(660..840)
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(855..1057)
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer complement(1058..1437)
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 1703..1807
/label="AmpR promoter"
CDS 1808..2665
/label="AmpR"
/note="beta-lactamase"
rep_origin 2839..3427
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3715..3736
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3751..3781
/label="lac promoter"
/note="promoter for the E. coli lac operon"
polyA_signal complement(3866..3999)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(4132..4503)
/label="BleoR"
/note="antibiotic-binding protein"
promoter complement(4522..4569)
/label="EM7 promoter"
/note="synthetic bacterial promoter"
promoter complement(4617..4946)
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin complement(4960..5388)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal complement(5434..5658)
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
LTR complement(5690..5870)
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter 6475..7653
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 7669..8055
/label="MS2-N55K"
/note="bacteriophage MS2 coat protein"
CDS 8146..8688
/label="p65"
/note="C-terminal portion of the p65 subunit of mouse
NF-kappa-B"
CDS 8713..9084
/label="HSF1"
/note="C-terminal activation domain from the human heat
shock transcription factor HSF1"
CDS 9100..9153
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 9157..10179
/label="HygR"
/note="aminoglycoside phosphotransferase from E. coli"
misc_feature 10216..10804
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
misc_feature complement(10857..10974)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(11082..11123)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(11272..11316)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
misc_feature complement(11501..11734)
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."