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.
- 载体名称:
- lentiGuide-Puro
- 载体抗性:
- Ampicillin
- 载体长度:
- 10183 bp
- 载体类型:
- CRISPR Plasmids
- 复制子:
- ori
- 载体来源:
- Sanjana NE, Shalem O, Zhang F.
- 拷贝数:
- High copy number
- 启动子:
- EF-1α
lentiGuide-Puro 载体载体图谱
lentiGuide-Puro 载体载体序列
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"
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