Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V012328 | lentiCRISPR v2 | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
- Vector Name:
- lentiCRISPR v2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 14873 bp
- Type:
- CRISPR Plasmids
- Replication origin:
- ori
- Source/Author:
- Sanjana NE, Shalem O, Zhang F.
- Copy Number:
- High copy number
- Promoter:
- U6
lentiCRISPR v2 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 ℃.
lentiCRISPR v2 vector Sequence
LOCUS lentiCRISPR_v2. 14873 bp DNA circular SYN 01-JAN-1980 DEFINITION Zhang lab lentiviral transfer plasmid, also known as pLentiCRISPR v2, for CRISPR/Cas9 genome editing of mammalian cells with a single guide RNA (sgRNA). ACCESSION . VERSION . KEYWORDS lentiCRISPR v2. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14873) AUTHORS Sanjana NE, Shalem O, Zhang F. TITLE Improved vectors and genome-wide libraries for CRISPR screening. JOURNAL Nat. Methods 2014;11:783-4. PUBMED 25075903 REFERENCE 2 (bases 1 to 14873) AUTHORS Zhang Lab / Addgene #52961 TITLE Direct Submission REFERENCE 3 (bases 1 to 14873) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Methods"; date: "2014"; volume: "11"; pages: "783-4" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT Full sequence confirmed by Addgene. FEATURES Location/Qualifiers source 1..14873 /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_feature 2853..4737 /label=filler /note="filler" /note="digest with BsmBI to replace the filler with an sgRNA " misc_RNA 4738..4813 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 4814..4819 /note="polIII terminator" /note="RNA polymerase III transcription terminator" 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 9215..9262 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=bipartite nuclear localization signal from nucl /note="nucleoplasmin NLS" /translation="KRPAATKKAGQAKKKK" CDS 9263..9286 /label=FLAG /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" CDS 9296..9352 /label=P2A /note="2A peptide from porcine teschovirus-1 polyprotein" CDS 9353..9946 /label=PuroR /note="puromycin N-acetyltransferase" CDS complement(10436..10447) /label=Factor Xa site /note="Factor Xa recognition and cleavage site" LTR 10625..10858 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 10890..11114 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 11160..11588 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 11602..11931 /label=SV40 promoter /note="SV40 enhancer and early promoter" promoter 11979..12026 /label=EM7 promoter /note="synthetic bacterial promoter" CDS 12045..12416 /label=BleoR /note="antibiotic-binding protein" polyA_signal 12549..12682 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(12767..12797) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(12812..12833) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(13121..13709) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(13883..14740) /label=AmpR /note="beta-lactamase" promoter complement(14741..14845) /label=AmpR promoter