Basic Vector Information
- Vector Name:
- pLV3-U6-MCS-shRNA-Fluc-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9234 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- hPGK
- 5' Primer:
- U6-F：GAGGGCCTATTTCCCATGA
- Growth Temperature:
- 37℃
pLV3-U6-MCS-shRNA-Fluc-Puro 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-MCS-shRNA-Fluc-Puro vector Sequence
LOCUS 62056_15715 9234 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9234)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9234
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 7..507
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 529..1125
/label=PuroR
/note="puromycin N-acetyltransferase"
LTR 1256..1489
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 1561..1695
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 1722..1857
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(1878..1896)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1906..1922)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2064..2519
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2545..2649
/label=AmpR promoter
CDS 2650..3507
/label=AmpR
/note="beta-lactamase"
rep_origin 3681..4269
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4557..4578
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4593..4623
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4631..4647
/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 4655..4671
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 4692..4710
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 4738..4964
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 4965..5145
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5192..5317
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5810..6043
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6228..6272
/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 6421..6462
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
promoter 6570..6810
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
enhancer 6832..7135
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 7136..7339
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 7374..9023
/label=luciferase
/note="firefly luciferase"
misc_feature 9065..9182
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 9231..9234
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
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