载体基本信息 | |||
载体名称 | eSpCas9(1.1) | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 8506 bp | 载体类型 | CRISPR Plasmids |
载体来源 | Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F. | 拷贝数 | High copy number |
eSpCas9(1.1) 载体载体图谱
eSpCas9(1.1) 载体载体序列
LOCUS eSpCas9(1.1). 8506 bp DNA circular SYN 01-JAN-1980 DEFINITION Zhang lab vector for expressing a single guide RNA (sgRNA) together with an S. pyogenes Cas9 variant with improved targeting specificity. ACCESSION . VERSION . KEYWORDS eSpCas9(1.1). SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8506) AUTHORS Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F. TITLE Rationally engineered Cas9 nucleases with improved specificity. JOURNAL Science 2016;351:84-8. PUBMED 26628643 REFERENCE 2 (bases 1 to 8506) AUTHORS Zhang Lab / Addgene #71814 TITLE Direct Submission REFERENCE 3 (bases 1 to 8506) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science"; date: "2016"; volume: "351"; pages: "84-8" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT An error in codon 848 was corrected based on sequencing results from Addgene. FEATURES Location/Qualifiers source 1..8506 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 268..343 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 344..349 /note="polIII terminator" /note="RNA polymerase III transcription terminator" enhancer 440..725 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 727..1004 /label=chicken beta-actin promoter intron 1005..1233 /note="hybrid intron" /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" CDS 1251..1253 /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" CDS 1254..1319 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=three tandem FLAG /note="3xFLAG" /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1326..1346 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /note="SV40 NLS" /translation="PKKKRKV" CDS 1371..5471 /label=eSpCas9(1.1) /note="Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system, mutated to improve targeting specificity (Slaymaker et al., 2016)" CDS 5472..5519 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=bipartite nuclear localization signal from nucl /note="nucleoplasmin NLS" /translation="KRPAATKKAGQAKKKK" polyA_signal 5553..5760 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 5769..5909 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 5984..6439 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 6721..6825 /label=AmpR promoter CDS 6826..7683 /label=AmpR /note="beta-lactamase" rep_origin 7857..8445 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
我们不提供该载体,载体信息仅供查询和参考。