Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011040 | pTRIP-SFFV-mtagBFP-2A STING | 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:
- pTRIP-SFFV-mtagBFP-2A STING
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11496 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- SFFV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- SFFV-F
pTRIP-SFFV-mtagBFP-2A STING 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.
pTRIP-SFFV-mtagBFP-2A STING vector Sequence
LOCUS V011040 11496 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V011040
VERSION V011040
KEYWORDS pTRIP-SFFV-mtagBFP-2A STING
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11496)
AUTHORS Cerboni S, Jeremiah N, Gentili M, Gehrmann U, Conrad C, Stolzenberg
MC, Picard C, Neven B, Fischer A, Amigorena S, Rieux-Laucat F, Manel
N
TITLE Intrinsic antiproliferative activity of the innate sensor STING in T
lymphocytes.
JOURNAL J Exp Med. 2017 Jun 5;214(6):1769-1785. doi: 10.1084/jem.20161674.
Epub 2017 May 8.
PUBMED 28484079
REFERENCE 2 (bases 1 to 11496)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11496)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1084/jem.20161674"; journalName: "J Exp Med"; date: "2017-06-5-
5"; volume: "214"; issue: "6"; pages: "1769-1785"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11496
/mol_type="other DNA"
/organism="synthetic DNA construct"
intron 43..108
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 238..258
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 683..817
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 985..1618
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 1665..1790
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2283..2516
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2701..2745
/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 2894..2935
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 3038..3155
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 3295..3702
/label="SFFV promoter"
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
CDS 3721..4419
/label="TagBFP"
/note="monomeric blue fluorescent protein"
CDS 4429..4485
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label="P2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 4492..5628
/gene="STING1"
/label="Stimulator of interferon genes protein"
/note="Stimulator of interferon genes protein from Homo
sapiens. Accession#: Q86WV6"
LTR 5900..6080
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(7608..7627)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(7841..7860)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7960..7982
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(8020..8038)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 8106..8210
/label="AmpR promoter"
CDS 8211..9068
/label="AmpR"
/note="beta-lactamase"
rep_origin 9242..9830
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9984..10001
/label="L4440"
/note="L4440 vector, forward primer"
promoter 10076..10405
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"