Basic Vector Information
- Vector Name:
- pLV2-U6-sgRNA-CMV-Cre
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10173 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- U6
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLV2-U6-sgRNA-CMV-Cre 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 ℃.
pLV2-U6-sgRNA-CMV-Cre vector Sequence
LOCUS V014145 10173 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014145
VERSION V014145
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10173)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10173
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 241..474
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 659..703
/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 852..893
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 1001..1118
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 1163..1183
/label="attB4"
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 1215..1455
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 1465..1494
/label="RfxCas13d DR30"
/note="5' processed 30-base direct repeat for an RfxCas13d
gRNA (Konermann et al., 2018)"
promoter 1880..2083
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 2130..2154
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2200..2220
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 2239..3267
/label="Cre"
/note="site-specific recombinase"
CDS 3286..3333
/label="nucleoplasmin NLS"
/note="bipartite nuclear localization signal from
nucleoplasmin"
protein_bind complement(3368..3392)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 3426..4014
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
polyA_signal 4751..4975
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 5021..5449
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5463..5792
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 5840..5887
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 5906..6277
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 6410..6543
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(6628..6658)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(6673..6694)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6982..7570)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7744..8601)
/label="AmpR"
/note="beta-lactamase"
promoter complement(8602..8706)
/label="AmpR promoter"
enhancer 8972..9351
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
LTR 9569..9749
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 9796..9921
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
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