Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001632 | pLKO.1.sh.beta-catenin.1248 | 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:
- pLKO.1.sh.beta-catenin.1248
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7083 bp
- Type:
- Mammalian Expression, Lentiviral, RNAi
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- hPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- LKO.1 5'
pLKO.1.sh.beta-catenin.1248 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.
pLKO.1.sh.beta-catenin.1248 vector Sequence
LOCUS V001632 7083 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001632
VERSION V001632
KEYWORDS pLKO.1.sh.beta-catenin.1248
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7083)
AUTHORS Firestein R, Bass AJ, Kim SY, Dunn IF, Silver SJ, Guney I, Freed E,
Ligon AH, Vena N, Ogino S, Chheda MG, Tamayo P, Finn S, Shrestha Y,
Boehm JS, Jain S, Bojarski E, Mermel C, Barretina J, Chan JA,
Baselga J, Tabernero J, Root DE, Fuchs CS, Loda M, Shivdasani RA,
Meyerson M, Hahn WC
TITLE CDK8 is a colorectal cancer oncogene that regulates beta-catenin
activity.
JOURNAL Nature. 2008 Sep 25. 455(7212):547-51.
PUBMED 18794900
REFERENCE 2 (bases 1 to 7083)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7083)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
2008 Sep 25. 455(7212):547-51."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7083
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 50..67
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 184..205
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 220..250
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 258..274
/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 282..298
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 319..337
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 365..591
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 592..772
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 819..944
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1437..1670
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1855..1899
/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 2048..2089
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter 2197..2437
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_feature 2541..2658
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2707..3217
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
CDS 3239..3835
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 3966..4199
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4271..4405
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4432..4567
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(4588..4606)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4616..4632)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4774..5229
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5255..5359
/label="AmpR promoter"
CDS 5360..6217
/label="AmpR"
/note="beta-lactamase"
rep_origin 6391..6979
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"