Basic Vector Information
- Vector Name:
- pLV3-U6-shRNA-EGFP-Neo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8312 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Neo/G418
- Promoter:
- SV40
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV3-U6-shRNA-EGFP-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 ℃.
pLV3-U6-shRNA-EGFP-Neo vector Sequence
LOCUS V014244 8312 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014244
VERSION V014244
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8312)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8312
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 59..388
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 397..1188
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase from Tn5"
LTR 1319..1552
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 1624..1758
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(1941..1959)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1969..1985)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2127..2582
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2608..2712
/label="AmpR promoter"
CDS 2713..3570
/label="AmpR"
/note="beta-lactamase"
rep_origin 3744..4332
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4620..4641
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4656..4686
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4694..4710
/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 4718..4734
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 4755..4773
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 4801..5027
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 5255..5380
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5873..6106
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6291..6335
/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 6484..6525
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter 6633..6873
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
enhancer 6895..7198
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 7199..7402
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 7437..8153
/label="EGFP"
/note="enhanced GFP"
misc_feature 8195..8312
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
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