Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010596 | pHR_PGK_antiHer24D5-7_synNotch_Gal4VP64 | 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:
- pHR_PGK_antiHer24D5-7_synNotch_Gal4VP64
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11423 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- mPGK
- Cloning Method:
- Gibson Cloning
- 5' Primer:
- gcttcaaaagcgcacgtct
pHR_PGK_antiHer24D5-7_synNotch_Gal4VP64 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.
pHR_PGK_antiHer24D5-7_synNotch_Gal4VP64 vector Sequence
LOCUS V010596 11423 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V010596
VERSION V010596
KEYWORDS pHR_PGK_antiHer24D5-7_synNotch_Gal4VP64
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11423)
AUTHORS Roybal KT, Williams JZ, Morsut L, Rupp LJ, Kolinko I, Choe JH,
Walker WJ, McNally KA, Lim WA
TITLE Engineering T Cells with Customized Therapeutic Response Programs
Using Synthetic Notch Receptors.
JOURNAL Cell. 2016 Oct 6;167(2):419-432.e16. doi:
10.1016/j.cell.2016.09.011. Epub 2016 Sep 29.
PUBMED 27693353
REFERENCE 2 (bases 1 to 11423)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11423)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.09.011"; journalName: "Cell"; date: "2016-10-6-
6"; volume: "167"; issue: "2"; pages: "419-432.e16"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11423
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 64..93
/label="Myc"
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 1810..2250
/label="GAL4 DNA binding domain"
/note="DNA binding domain of the GAL4 transcriptional
activator"
CDS 2287..2436
/label="VP64"
/note="tetrameric repeat of the minimal activation domain
of herpes simplex virus VP16 (Beerli et al., 1998)"
misc_feature 2555..3141
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 3206..3439
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(3464..3482)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3772..3791)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(4005..4024)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4124..4146
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(4184..4202)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4270..4374
/label="AmpR promoter"
CDS 4375..5232
/label="AmpR"
/note="beta-lactamase"
rep_origin 5406..5994
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6148..6165
/label="L4440"
/note="L4440 vector, forward primer"
promoter 6240..6569
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 7703..7768
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 7898..7918
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 8343..8477
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 8645..9278
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 9325..9450
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 9947..10180
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 10365..10409
/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 10558..10599
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 10694..10811
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 10871..11370
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"