Basic Vector Information
- Vector Name:
- pLV3-EF1a-dCas9-VPR-EGFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 15336 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- EF-1α
- Growth Temperature:
- 30℃
pLV3-EF1a-dCas9-VPR-EGFP 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-EF1a-dCas9-VPR-EGFP vector Sequence
LOCUS 62056_15490 15336 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 15336)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..15336
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 618..820
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1680..1913
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2098..2142
/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 2291..2332
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2440..2557
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2687..3865
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3884..3904
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 3914..8017
/label=Cas9m4
/note="catalytically dead mutant of the Cas9 endonuclease
from the Streptococcus pyogenes Type II CRISPR/Cas system"
CDS 8072..8092
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 9287..10000
/label=EGFP
/note="enhanced GFP"
misc_feature 10028..10616
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 11141..11321
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 11353..11577
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 11623..12051
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 12065..12394
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 12442..12489
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 12508..12879
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 13012..13145
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(13230..13260)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(13275..13296)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(13584..14172)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(14346..15203)
/label=AmpR
/note="beta-lactamase"
promoter complement(15204..15308)
/label=AmpR promoter
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