Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007219 | pInducer20 | 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:
- pInducer20
- Antibiotic Resistance:
- Chloramphenicol, Ampicillin
- Length:
- 11808 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Promoter:
- UbC
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- LNCX_primer
pInducer20 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.
pInducer20 vector Sequence
LOCUS V007219 11808 bp DNA circular SYN 08-SEP-2021
DEFINITION Exported.
ACCESSION V007219
VERSION V007219
KEYWORDS pInducer20
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11808)
AUTHORS Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE,
Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi
RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF,
Elledge SJ
TITLE The pINDUCER lentiviral toolkit for inducible RNA interference in
vitro and in vivo.
JOURNAL Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi:
10.1073/pnas.1019736108. Epub 2011 Feb 9.
PUBMED 21307310
REFERENCE 2 (bases 1 to 11808)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11808)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1073/pnas.1019736108"; journalName: "Proc Natl Acad Sci U S A";
date: "2011-03-1- 1"; volume: "108"; issue: "9"; pages: "3665-70"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11808
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(75..818)
/label="rtTA-Advanced"
/note="improved tetracycline-controlled transactivator"
primer_bind complement(1021..1040)
/label="hUBCpro-F"
/note="Human Ubiquitin C (UbC) promoter, forward primer"
misc_feature complement(1255..1811)
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS complement(2174..2200)
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
protein_bind 2253..2377
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(2421..2723)
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS complement(3068..3724)
/label="CmR"
/note="chloramphenicol acetyltransferase"
promoter complement(3778..3808)
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
protein_bind complement(3833..3957)
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(4066..4090)
/label="LNCX"
/note="Human CMV promoter, forward primer"
promoter complement(4087..4125)
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind complement(4158..4428)
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
misc_feature complement(4483..4600)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(4703..4744)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(4893..4937)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
misc_feature complement(5122..5355)
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature complement(5852..5977)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(6024..6657)
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6849..6868)
/label="EBV-rev"
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6918..6934)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6918..6934)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6931..6953)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 6942..6958
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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(6966..6996)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(7011..7032)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7161..7180)
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
primer_bind complement(7272..7289)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(7443..8031)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8205..9062)
/label="AmpR"
/note="beta-lactamase"
promoter complement(9063..9167)
/label="AmpR promoter"
primer_bind 9235..9253
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
LTR complement(9372..9605)
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature complement(9680..10268)
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(10448..11239)
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
misc_feature complement(11255..11805)
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"