Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012530 | pTRIPZ empty | 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
The Thermo Scientific TRIPZ Inducible Lentiviral shRNA Collection was developed in collaboration with Dr. Greg Hannon of Cold Spring Harbor Laboratory (CSHL) and Dr. Steve Elledge of Harvard Medical School. This collection combines the design advantages of microRNA-adapted short hairpin RNA (shRNA) with the pTRIPZ lentiviral inducible vector to create a powerful RNAi trigger capable of producing gene silencing. The vector is engineered to be Tet-On and produces tightly regulated induction of shRNA expression in the presence of doxycycline.
- Vector Name:
- pTRIPZ empty
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13320 bp
- Type:
- Lentiviral vectors
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High copy number
- Promoter:
- UbC
- Cloning Method:
- Enzyme digestion and ligation
pTRIPZ empty 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.
pTRIPZ empty vector Sequence
LOCUS V012530 13320 bp DNA circular SYN 13-JAN-2022
DEFINITION Exported.
ACCESSION V012530
VERSION V012530
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13320)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 13320)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13320
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/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 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2063..2180
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(2248..2619)
/label="BleoR"
/note="antibiotic-binding protein"
promoter complement(2639..2686)
/label="EM7 promoter"
/note="synthetic bacterial promoter"
protein_bind 2722..2740
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2757..2775
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2793..2811
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2829..2847
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2864..2882
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2900..2918
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 2930..2968
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 3005..3697
/label="TurboRFP"
/note="red fluorescent protein from Entacmaea quadricolor"
ncRNA 3713..3806
/label="5' miR-30a"
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
ncRNA 3832..3944
/label="3' miR-30a"
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
promoter complement(4039..4057)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 4070..5281
/label="UbC promoter"
/note="human ubiquitin C promoter"
protein_bind 5294..5327
/label="Cre recombinase binding site"
/bound_moiety="Cre recombinase"
/note="loxP511"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GTATACAT). loxP511 sites are compatible with each
other, but incompatible with wild-type loxP sites."
CDS 5334..6038
/label="rtTA3"
/note="reverse tetracycline transactivator 3"
protein_bind complement(6042..6075)
/label="loxP511"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are
compatible with each other, but incompatible with wild-type
loxP sites."
misc_feature 6098..6680
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6679..7272
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 7291..7879
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 8086..8319
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 8426..8650
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 8696..9124
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 9138..9467
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
polyA_signal 10675..10796
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(10845..10861)
/label="M13 rev"
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 10869..10885
/label="lac repressor encoded by lacI binding site"
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
promoter complement(10893..10923)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(10938..10959)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(11247..11835)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12009..12866)
/label="AmpR"
/note="beta-lactamase"
promoter complement(12867..12971)
/label="AmpR promoter"
polyA_signal 13019..13153
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"