Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011344 | pINDUCER21 (ORF-EG) | 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:
- pINDUCER21 (ORF-EG)
- Antibiotic Resistance:
- Chloramphenicol, Ampicillin
- Length:
- 12152 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Selection Marker:
- eGFP
- Promoter:
- EF-1α
pINDUCER21 (ORF-EG) 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.
pINDUCER21 (ORF-EG) vector Sequence
LOCUS V011344 12152 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V011344
VERSION V011344
KEYWORDS pINDUCER21 (ORF-EG)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12152)
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 12152)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12152)
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..12152
/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 2058..2175
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2230..2500
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2533..2571
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2568..2592
/label="LNCX"
/note="Human CMV promoter, forward primer"
protein_bind 2701..2825
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 2850..2880
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
CDS 2934..3590
/label="CmR"
/note="chloramphenicol acetyltransferase"
CDS 3935..4237
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(4281..4405)
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 4458..4484
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
promoter 4704..5882
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
protein_bind 5952..5985
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS 5992..6735
/label="rtTA-Advanced"
/note="improved tetracycline-controlled transactivator"
misc_feature 6791..7364
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 7380..8096
/label="EGFP"
/note="enhanced GFP"
primer_bind 8235..8254
/label="Neo-F"
/note="Neomycin resistance gene, forward primer"
misc_feature 8542..9130
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 9205..9438
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(9557..9575)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 9643..9747
/label="AmpR promoter"
CDS 9748..10605
/label="AmpR"
/note="beta-lactamase"
rep_origin 10779..11367
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11521..11538
/label="L4440"
/note="L4440 vector, forward primer"
primer_bind 11630..11649
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
protein_bind 11778..11799
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11814..11844
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 11852..11868
/label="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)."
primer_bind 11876..11892
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 11942..11961
/label="EBV-rev"
/note="SV40 polyA terminator, reverse primer"