Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V012327 | lentiGuide-Puro | 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
The lentiGuide-Puro is a lentiviral vector, also known as pLentiGuide-Puro. It is for expressing a single guide RNA (sgRNA) in Cas9-expressing cells.
lentiGuide-Puro 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 ℃.
References
- Datlinger P, Rendeiro AF, Schmidl C, Krausgruber T, Traxler P, Klughammer J, Schuster LC, Kuchler A, Alpar D, Bock C. Pooled CRISPR screening with single-cell transcriptome readout. Nat Methods. 2017 Mar;14(3):297-301.
lentiGuide-Puro vector Sequence
LOCUS lentiGuide-Puro. 10183 bp DNA circular SYN 01-JAN-1980 DEFINITION Zhang lab lentiviral vector, also known as pLentiGuide-Puro, for expressing a single guide RNA (sgRNA) in Cas9-expressing cells. Use together with lentiCas9-Blast. ACCESSION . VERSION . KEYWORDS lentiGuide-Puro. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10183) 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 10183) AUTHORS Zhang Lab / Addgene #52963 TITLE Direct Submission REFERENCE 3 (bases 1 to 10183) 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" FEATURES Location/Qualifiers source 1..10183 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 87..1265 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" CDS 1278..1874 /label=PuroR /note="puromycin N-acetyltransferase" misc_feature 1893..2481 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 2553..2786 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2864..2998 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 3025..3160 /label=SV40 ori /note="SV40 origin of replication" promoter complement(3181..3199) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" rep_origin 3367..3822 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3848..3952 /label=AmpR promoter CDS 3953..4810 /label=AmpR /note="beta-lactamase" rep_origin 4984..5572 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 5860..5881 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 5896..5926 /label=lac promoter /note="promoter for the E. coli lac operon" promoter 5995..6013 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6041..6267 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6268..6448 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6495..6620 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7113..7346 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7531..7575 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" promoter 7753..7993 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_feature 7998..9882 /label=filler /note="filler" /note="digest with BsmBI to replace the filler with an sgRNA" misc_RNA 9883..9958 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 9959..9964 /note="polIII terminator" /note="RNA polymerase III transcription terminator" misc_feature 10014..10131 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1"