Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000016 | Tet-pLKO-puro-Scrambled | 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
- Vector Name:
- Tet-pLKO-puro-Scrambled
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8815 bp
- Type:
- Mammalian Expression, Lentiviral, RNAi
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- hPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- H1
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
Tet-pLKO-puro-Scrambled 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.
Tet-pLKO-puro-Scrambled vector Sequence
LOCUS V000016 8815 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000016
VERSION V000016
KEYWORDS Tet-pLKO-puro-Scrambled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8815)
AUTHORS Rudin CM, Durinck S, Stawiski EW, Poirier JT, Modrusan Z, Shames DS,
Bergbower EA, Guan Y, Shin J, Guillory J, Rivers CS, Foo CK, Bhatt
D, Stinson J, Gnad F, Haverty PM, Gentleman R, Chaudhuri S,
Janakiraman V, Jaiswal BS, Parikh C, Yuan W, Zhang Z, Koeppen H, Wu
TD, Stern HM, Yauch RL, Huffman KE, Paskulin DD, Illei PB,
Varella-Garcia M, Gazdar AF, de Sauvage FJ, Bourgon R, Minna JD,
Brock MV, Seshagiri S
TITLE Comprehensive genomic analysis identifies SOX2 as a frequently
amplified gene in small-cell lung cancer.
JOURNAL Nat Genet. 2012 Oct;44(10):1111-6. doi: 10.1038/ng.2405. Epub 2012
Sep 2.
PUBMED 22941189
REFERENCE 2 (bases 1 to 8815)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8815)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ng.2405";
journalName: "Nat Genet"; date: "2012-10"; volume: "44"; issue:
"10"; pages: "1111-6"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8815
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 489..722
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 794..928
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 955..1090
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(1111..1129)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1139..1155)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 1297..1752
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1778..1882
/label="AmpR promoter"
CDS 1883..2740
/label="AmpR"
/note="beta-lactamase"
rep_origin 2914..3502
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3790..3811
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3826..3856
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3864..3880
/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 3888..3904
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3925..3943
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 3971..4197
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 4425..4550
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5043..5276
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5461..5505
/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 5654..5695
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter 5794..5888
/label="H1-2O2 promoter"
/note="doxycycline-inducible variant of the human H1 RNA
promoter (Henriksen et al., 2007)"
misc_feature 5988..6105
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 6154..6664
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
intron 6673..7245
/label="beta-globin intron"
/note="intron from rabbit beta-globin gene"
promoter 7300..7318
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 7329..7949
/label="TetR"
/note="tetracycline repressor TetR"
misc_feature 7984..8557
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS join(8577..8815,1..358)
/label="PuroR"
/note="puromycin N-acetyltransferase"