载体基本信息 | |||
载体名称 | lentiCas9-EGFP | 载体抗性 | Ampicillin |
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载体长度 | 13177 bp | 载体类型 | CRISPR Plasmids |
载体来源 | Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song | 拷贝数 | High copy number |
lentiCas9-EGFP 载体载体图谱
lentiCas9-EGFP 载体载体序列
LOCUS lentiCas9-EGFP. 13177 bp DNA circular SYN 01-JAN-1980 DEFINITION Zhang lab lentiviral vector for expressing human codon-optimized S. pyogenes Cas9 plus enhanced GFP (EGFP). ACCESSION . VERSION . KEYWORDS lentiCas9-EGFP. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13177) AUTHORS Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song J, Pan JQ, Weissleder R, Lee H, Zhang F, Sharp PA. TITLE Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis. JOURNAL Cell 2015;160:1246-60. PUBMED 25748654 REFERENCE 2 (bases 1 to 13177) AUTHORS TITLE Direct Submission REFERENCE 3 (bases 1 to 13177) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; date: "2015"; volume: "160"; pages: "1246-60" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..13177 /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 2650..2861 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" CDS 2886..6989 /label=Cas9 /note="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" CDS 6990..7037 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=bipartite nuclear localization signal from nucl /note="nucleoplasmin NLS" /translation="KRPAATKKAGQAKKKK" CDS 7038..7061 /label=FLAG /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" CDS 7071..7127 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=2A peptide from porcine teschovirus-1 polyprotein /note="P2A" /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 7128..7841 /label=EGFP /note="enhanced GFP" misc_feature 7869..8457 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 8982..9162 /label=5' LTR (truncated) /note="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 9194..9418 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 9464..9892 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 10017..10227 /label=SV40 promoter /note="SV40 early promoter" promoter 10283..10330 /label=EM7 promoter /note="synthetic bacterial promoter" CDS 10349..10720 /label=BleoR /note="antibiotic-binding protein" polyA_signal 10853..10986 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(11071..11101) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(11116..11137) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(11425..12013) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(12187..13044) /label=AmpR /note="beta-lactamase" promoter complement(13045..13149) /label=AmpR promoter
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