Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V016793 | pINDUCER11(miR-RUG) | 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:
- pINDUCER11(miR-RUG)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 14701 bp
- Type:
- RNA interference, Tetracycline inducible
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- UbC
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pINDUCER11(miR-RUG) 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.
pINDUCER11(miR-RUG) vector Sequence
LOCUS V016793 14701 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016793
VERSION V016793
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 14701)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..14701
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 500..733
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 918..962
/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 1111..1152
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 1260..1377
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(1445..1816)
/label="BleoR"
/note="antibiotic-binding protein"
promoter complement(1836..1883)
/label="EM7 promoter"
/note="synthetic bacterial promoter"
protein_bind 1953..2223
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2256..2294
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2391..3083
/label="TurboRFP"
/note="red fluorescent protein from Entacmaea quadricolor"
ncRNA 3137..3230
/label="5' miR-30a"
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
CDS 3268..3285
/label="thrombin site"
/note="thrombin recognition and cleavage site"
CDS 3286..3309
/label="HRV 3C site"
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
CDS complement(3486..4466)
/gene="pheS"
/label="Phenylalanine--tRNA ligase alpha subunit"
/note="Phenylalanine--tRNA ligase alpha subunit from
Escherichia coli O139:H28 (strain E24377A / ETEC).
Accession#: A7ZMI2"
CDS 4604..4624
/label="TEV site"
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
ncRNA 4648..4760
/label="3' miR-30a"
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
promoter 4811..6022
/label="UbC promoter"
/note="human ubiquitin C promoter"
protein_bind 6029..6062
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS 6069..6812
/label="rtTA-Advanced"
/note="improved tetracycline-controlled transactivator"
misc_feature 6864..7436
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 7454..8170
/label="EGFP"
/note="enhanced GFP"
misc_feature 8195..8783
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 8990..9223
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 9330..9554
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 9600..9980
/label="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 10042..10371
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
polyA_signal 11194..11275
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(11412..11442)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(11457..11478)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(11766..12354)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12528..13385)
/label="AmpR"
/note="beta-lactamase"
promoter complement(13386..13490)
/label="AmpR promoter"
polyA_signal 13538..13672
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 13840..14473
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 14520..14645
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"