| 载体基本信息 | |||
| 载体名称 | lenti dCAS-VP64_Blast | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 14085 bp | 载体类型 | CRISPR Plasmids |
| 载体来源 | Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena | 拷贝数 | High copy number |
lenti dCAS-VP64_Blast 载体载体图谱
lenti dCAS-VP64_Blast 载体载体序列
LOCUS lenti_dCAS-VP64_ 14085 bp DNA circular SYN 01-JAN-1980
DEFINITION Zhang lab lentiviral vector encoding a blasticidin resistance marker
plus catalytically inactive dCas9 fused to the VP64 transcriptional
activation domain.
ACCESSION .
VERSION .
KEYWORDS lenti dCAS-VP64_Blast.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14085)
AUTHORS Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F.
TITLE Genome-scale transcriptional activation by an engineered CRISPR-Cas9
complex.
JOURNAL Nature 2015;517:583-8.
PUBMED 25494202
REFERENCE 2 (bases 1 to 14085)
AUTHORS Zhang Lab / Addgene #61425
TITLE Direct Submission
REFERENCE 3 (bases 1 to 14085)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature";
date: "2015"; volume: "517"; pages: "583-8"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14085
/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 2687..3865
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3878..3880
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 3881..3928
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=bipartite nuclear localization signal from
nucl
/note="nucleoplasmin NLS"
/translation="KRPAATKKAGQAKKKK"
CDS 3929..8029
/label=dCas9
/note="catalytically dead mutant of the Cas9 endonuclease
from the Streptococcus pyogenes Type II CRISPR/Cas system"
CDS 8084..8104
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 8129..8278
/label=VP64
/note="tetrameric repeat of the minimal activation domain
of herpes simplex virus VP16 (Beerli et al., 1998)"
CDS 8300..8353
/label=T2A
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 8354..8749
/label=BSD
/note="blasticidin S deaminase"
misc_feature 8777..9365
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 9890..10070
/label=LTR (truncated)
/note="LTR (truncated)"
/note="truncated long terminal repeat (LTR) from HIV-1"
polyA_signal 10102..10326
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 10372..10800
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 10814..11143
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 11191..11238
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 11257..11628
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 11761..11894
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(11979..12009)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(12024..12045)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(12333..12921)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(13095..13952)
/label=AmpR
/note="beta-lactamase"
promoter complement(13953..14057)
/label=AmpR promoter
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