Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001414 | pHR_EGFPligand | 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_EGFPligand
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9979 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- SFFV
pHR_EGFPligand 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_EGFPligand vector Sequence
LOCUS V001414 9979 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001414
VERSION V001414
KEYWORDS pHR_EGFPligand
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9979)
AUTHORS Morsut L, Roybal KT, Xiong X, Gordley RM, Coyle SM, Thomson M, Lim
WA
TITLE Engineering Customized Cell Sensing and Response Behaviors Using
Synthetic Notch Receptors.
JOURNAL Cell. 2016 Feb 11;164(4):780-91. doi: 10.1016/j.cell.2016.01.012.
Epub 2016 Jan 28.
PUBMED 26830878
REFERENCE 2 (bases 1 to 9979)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9979)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.01.012"; journalName: "Cell"; date:
"2016-02-11- 11"; volume: "164"; issue: "4"; pages: "780-91"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9979
/mol_type="other DNA"
/organism="synthetic DNA construct"
intron 655..720
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 850..870
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 1295..1429
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 1597..2230
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 2277..2402
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2899..3132
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 3317..3361
/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 3510..3551
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 3646..3763
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 3916..4323
/label="SFFV promoter"
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 4391..4400
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4460..5176
/label="EGFP"
/note="enhanced GFP"
misc_feature 5439..6027
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6030..6046)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6031..6047)
/label="pBluescriptKS"
/note="For pBluescript vector"
LTR 6137..6370
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(6395..6413)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(6703..6722)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(6936..6955)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7055..7077
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(7115..7133)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7201..7305
/label="AmpR promoter"
CDS 7306..8163
/label="AmpR"
/note="beta-lactamase"
rep_origin 8337..8925
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9079..9096
/label="L4440"
/note="L4440 vector, forward primer"
promoter 9171..9500
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"