Basic Vector Information
- Vector Name:
- pLV3-U6-CasRx-pre-gRNA-CMV-Cre-NLS
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8084 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- U6
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV3-U6-CasRx-pre-gRNA-CMV-Cre-NLS 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-CasRx-pre-gRNA-CMV-Cre-NLS vector Sequence
LOCUS 62056_15625 8084 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 8084)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8084
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 6..233
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 458..583
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1076..1309
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1493..1537
/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 1686..1727
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 1804..1921
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1973..2213
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 2223..2258
/label=RfxCas13d DR36
/note="36-base direct repeat for an RfxCas13d pre-gRNA
(Konermann et al., 2018)"
enhancer 2356..2659
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2660..2863
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 2910..2934
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 2980..3000
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 3019..4047
/label=Cre
/note="site-specific recombinase"
CDS 4066..4113
/label=nucleoplasmin NLS
/note="bipartite nuclear localization signal from
nucleoplasmin"
protein_bind complement(4142..4166)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 4182..4770
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4841..5074
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5146..5280
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 5286..5421
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5459..5475)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5483..5499)
/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)."
promoter complement(5507..5537)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5552..5573)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5861..6449)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6623..7480)
/label=AmpR
/note="beta-lactamase"
promoter complement(7481..7585)
/label=AmpR promoter
primer_bind 8059..8075
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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