Basic Vector Information
- Vector Name:
- pLV3-U6-Negative-Control-shRNA-mCherry-Neo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8053 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Neo/G418
- Promoter:
- hPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV3-U6-Negative-Control-shRNA-mCherry-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-Negative-Control-shRNA-mCherry-Neo vector Sequence
LOCUS V014215 8053 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014215
VERSION V014215
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8053)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8053
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 7..507
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
CDS 516..1223
/label="mCherry"
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
CDS 1299..2090
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase from Tn5"
LTR 2221..2454
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2526..2660
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 2687..2822
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(2843..2861)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2871..2887)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3029..3484
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3510..3614
/label="AmpR promoter"
CDS 3615..4472
/label="AmpR"
/note="beta-lactamase"
rep_origin 4646..5234
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5522..5543
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5558..5588
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5596..5612
/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 5620..5636
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 5657..5675
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 5703..5929
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 5930..6110
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6157..6282
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6775..7008
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7193..7237
/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 7386..7427
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter 7535..7775
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_feature 7884..8001
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 8050..8053
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
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