Basic Vector Information
3rd generation lentiviral plasmid for inducible expression of shRNA; neomycin selection.
- Vector Name:
- Tet-pLKO-neo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10959 bp
- Type:
- RNAi
- Replication origin:
- ori
- Source/Author:
- Dmitri Wiederschain
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- hPGK
Tet-pLKO-neo vector Vector Map
Plasmid Resuspension 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 ℃.
Tet-pLKO-neo vector Sequence
LOCUS Tet-pLKO-neo. 10959 bp DNA circular SYN 11-SEP-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS Tet-pLKO-neo.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10959)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 10959)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10959
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 106..210
/label=AmpR promoter
CDS 211..1068
/label=AmpR
/note="beta-lactamase"
rep_origin 1242..1830
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2118..2139
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2154..2184
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2192..2208
/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)."
primer_bind 2216..2232
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 2253..2271
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2299..2525
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 2526..2706
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2753..2878
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3371..3604
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 3789..3833
/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 3982..4023
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
promoter 4122..4216
/label=H1-2O2 promoter
/note="doxycycline-inducible variant of the human H1 RNA
promoter (Henriksen et al., 2007)"
misc_feature 6134..6251
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 6300..6810
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
intron 6819..7391
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
promoter 7446..7464
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 7475..8095
/label=TetR
/note="tetracycline repressor TetR"
misc_feature 8130..8703
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 8710..9501
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
protein_bind complement(9533..9632)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
LTR 9776..10009
/label=3' LTR (Delta-U3)
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 10081..10215
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 10242..10377
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(10398..10416)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(10426..10442)
/label=M13 fwd
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 10584..10959
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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