Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012333 | lenti MS2-P65-HSF1_Hygro | 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:
- lenti MS2-P65-HSF1_Hygro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11736 bp
- Type:
- CRISPR Plasmids
- Replication origin:
- ori
- Source/Author:
- Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
- Copy Number:
- High copy number
- Promoter:
- EF-1α
lenti MS2-P65-HSF1_Hygro 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.
lenti MS2-P65-HSF1_Hygro vector Sequence
LOCUS V012333 11736 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V012333
VERSION V012333
KEYWORDS lenti MS2-P65-HSF1_Hygro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11736)
AUTHORS Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F.
TITLE Genome-scale transcriptional activation by an engineered CRISPR-Cas9
complex.
JOURNAL Nature 2015;517:583-8.
PUBMED 25494202
REFERENCE 2 (bases 1 to 11736)
AUTHORS Zhang Lab / Addgene #61426
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11736)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature";
date: "2015"; volume: "517"; pages: "583-8"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11736
/mol_type="other DNA"
/organism="synthetic DNA construct"
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 1680..1913
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2098..2142
/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 2291..2332
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2440..2557
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2687..3865
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3878..3880
/codon_start=1
/product="start codon"
/label="start codon"
/note="ATG"
/translation="M"
CDS 3881..4267
/label="MS2-N55K"
/note="bacteriophage MS2 coat protein"
CDS 4322..4342
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
CDS 4358..4900
/label="p65"
/note="C-terminal portion of the p65 subunit of mouse
NF-kappa-B"
CDS 4925..5296
/label="HSF1"
/note="C-terminal activation domain from the human heat
shock transcription factor HSF1"
CDS 5312..5365
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 5369..6391
/label="HygR"
/note="aminoglycoside phosphotransferase from E. coli"
misc_feature 6428..7016
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 7541..7721
/label="5' LTR (truncated)"
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 7753..7977
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 8023..8451
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8465..8794
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 8842..8889
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 8908..9279
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 9412..9545
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(9630..9660)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(9675..9696)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(9984..10572)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10746..11603)
/label="AmpR"
/note="beta-lactamase"
promoter complement(11604..11708)
/label="AmpR promoter"