载体基本信息 | |||
载体名称 | lentiCas9n(D10A)-Blast | 载体抗性 | Ampicillin |
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载体长度 | 12860 bp | 载体类型 | CRISPR Plasmids |
载体来源 | Sanjana NE, Shalem O, Zhang F. | 拷贝数 | High copy number |
lentiCas9n(D10A)-Blast 载体载体图谱
lentiCas9n(D10A)-Blast 载体载体序列
LOCUS lentiCas9n(D10A) 12860 bp DNA circular SYN 01-JAN-1980 DEFINITION Zhang lab lentiviral vector with a blasticidin resistance marker, for expressing Cas9 with the D10A nickase mutation. ACCESSION . VERSION . KEYWORDS lentiCas9n(D10A)-Blast. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12860) 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 12860) AUTHORS Zhang Lab / Addgene #63593 TITLE Direct Submission REFERENCE 3 (bases 1 to 12860) 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..12860 /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 2651..2862 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" CDS 2887..6990 /label=Cas9(D10A) /note="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" CDS 6991..7038 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=bipartite nuclear localization signal from nucl /note="nucleoplasmin NLS" /translation="KRPAATKKAGQAKKKK" CDS 7039..7062 /label=FLAG /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" CDS 7072..7128 /label=P2A /note="2A peptide from porcine teschovirus-1 polyprotein" CDS 7129..7524 /label=BSD /note="blasticidin S deaminase" misc_feature 7552..8140 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 8665..8845 /label=5' LTR (truncated) /note="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 8877..9101 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 9147..9575 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 9589..9918 /label=SV40 promoter /note="SV40 enhancer and early promoter" promoter 9966..10013 /note="EM7 promoter" /note="synthetic bacterial promoter " CDS 10032..10403 /label=BleoR /note="antibiotic-binding protein" polyA_signal 10536..10669 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(10754..10784) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(10799..10820) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(11108..11696) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(11870..12727) /label=AmpR /note="beta-lactamase" promoter complement(12728..12832) /label=AmpR promoter
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