- 载体名称:
- eSpCas9(1.1)
- 载体抗性:
- Ampicillin
- 载体长度:
- 8506 bp
- 载体类型:
- CRISPR Plasmids
- 复制子:
- ori
- 载体来源:
- Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F.
- 拷贝数:
- High copy number
- 启动子:
- CBh
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。