Basic Vector Information
- Vector Name:
- pLV3-U6-MCS-sgRNA-Cas9-Fluc-Blast
- Antibiotic Resistance:
- Ampicillin
- Length:
- 16388 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Blast
- Promoter:
- SV40
- 5' Primer:
- U6-F：GAGGGCCTATTTCCCATGA
- Growth Temperature:
- 37℃
pLV3-U6-MCS-sgRNA-Cas9-Fluc-Blast 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-sgRNA-Cas9-Fluc-Blast vector Sequence
LOCUS 62056_15650 16388 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 16388)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..16388
/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 2608..2848
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 4738..4813
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 4875..5086
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 5111..9214
/label=Cas9
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
CDS 9263..9286
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 9293..9349
/label=P2A
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 9350..10996
/label=luciferase
/note="firefly luciferase"
CDS 11057..11452
/label=BSD
/note="blasticidin S deaminase"
misc_feature 11480..12068
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 12140..12373
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 12405..12629
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 12675..13103
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 13117..13446
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 13494..13541
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 13560..13931
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 14064..14197
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(14282..14312)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(14327..14348)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(14636..15224)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(15398..16255)
/label=AmpR
/note="beta-lactamase"
promoter complement(16256..16360)
/label=AmpR promoter
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