Basic Vector Information
- Vector Name:
- pLV3-U6-ccdB-shRNA-hPGK-Fluc-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9555 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- hPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV3-U6-ccdB-shRNA-hPGK-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-ccdB-shRNA-hPGK-Fluc-Puro vector Sequence
LOCUS V014295 9555 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014295
VERSION V014295
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9555)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9555
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 7..507
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
CDS 529..2178
/label="luciferase"
/note="firefly luciferase"
CDS 2179..2244
/label="F2A"
/note="2A peptide from foot-and-mouth disease virus
polyprotein"
CDS 2245..2841
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 2972..3205
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 3277..3411
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 3438..3573
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(3594..3612)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3622..3638)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3780..4235
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4261..4365
/label="AmpR promoter"
CDS 4366..5223
/label="AmpR"
/note="beta-lactamase"
rep_origin 5397..5985
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6273..6294
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6309..6339
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6347..6363
/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 6371..6387
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 6408..6426
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6454..6680
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 6681..6861
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6908..7033
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7526..7759
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7944..7988
/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 8137..8178
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter 8286..8526
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
CDS complement(8595..8897)
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
promoter complement(9249..9279)
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
misc_feature 9386..9503
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 9552..9555
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
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