Basic Vector Information
- Vector Name:
- pLVPT-tTR-KRAB
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11634 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- mPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- See map
pLVPT-tTR-KRAB 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 ℃.
pLVPT-tTR-KRAB vector Sequence
LOCUS V006525 11634 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006525
VERSION V006525
KEYWORDS pLVPT-tTR-KRAB.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11634)
AUTHORS Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P
TITLE A versatile tool for conditional gene expression and knockdown.
JOURNAL Nat Methods. 2006 Feb . 3(2):109-16.
PUBMED 16432520
REFERENCE 2 (bases 1 to 11634)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11634)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2006 Feb . 3(2):109-16."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11634
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 176..217
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 294..411
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 450..949
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 1006..1722
/label="EGFP"
/note="enhanced GFP"
misc_feature 1805..2367
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 2372..2992
/label="TetR"
/note="tetracycline repressor TetR"
CDS 2993..3013
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 3116..3310
/label="KRAB"
/note="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
misc_feature 3480..4068
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
protein_bind 4206..4476
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
LTR 4515..4695
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(4720..4738)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 5526..5630
/label="AmpR promoter"
CDS 5631..6488
/label="AmpR"
/note="beta-lactamase"
rep_origin 6662..7250
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 7496..7825
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 8959..9024
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 9154..9174
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 9599..9733
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 10277..10534
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 10581..10706
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 11199..11432
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS join(11617..11634,1..27)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
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