Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007496 | pHR-tdPP7-3xmCherry | 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-tdPP7-3xmCherry
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11752 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- SFFV
pHR-tdPP7-3xmCherry 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.
pHR-tdPP7-3xmCherry vector Sequence
LOCUS V007496 11752 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007496
VERSION V007496
KEYWORDS pHR-tdPP7-3xmCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11752)
AUTHORS Yan X, Hoek TA, Vale RD, Tanenbaum ME
TITLE Dynamics of Translation of Single mRNA Molecules In Vivo.
JOURNAL Cell. 2016 May 5;165(4):976-89. doi: 10.1016/j.cell.2016.04.034.
PUBMED 27153498
REFERENCE 2 (bases 1 to 11752)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11752)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.04"; journalName: "Cell"; date: "2016-05-5- 5";
volume: "165"; issue: "4"; pages: "976-89"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11752
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(302..321)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 655..677
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(715..733)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 801..905
/label="AmpR promoter"
CDS 906..1763
/label="AmpR"
/note="beta-lactamase"
rep_origin 1937..2525
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2679..2696
/label="L4440"
/note="L4440 vector, forward primer"
promoter 2771..3100
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 4234..4299
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4429..4449
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 4874..5008
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 5177..5810
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 5857..5982
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6474..6707
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6892..6936
/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 7085..7126
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 7221..7338
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 7491..7898
/label="SFFV promoter"
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
CDS 7945..8328
/note="Capsid protein from Pseudomonas phage PP7.
Accession#: P03630"
/label="Capsid protein"
CDS 8797..9501
/label="mCherry"
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
CDS 9547..9708
/codon_start=1
/product="B1 domain of Streptococcal protein G "
/label="GB1"
/note="effective as a solubilizing fusion partner (Cheng
and Patel, 2004)"
/translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
ATKTFTVTE"
CDS 9733..10437
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label="mCherry"
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
CDS 10483..10644
/codon_start=1
/product="B1 domain of Streptococcal protein G "
/label="GB1"
/note="effective as a solubilizing fusion partner (Cheng
and Patel, 2004)"
/translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
ATKTFTVTE"
CDS 10702..11406
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label="mCherry"
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
LTR 11488..11721
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"