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Plasmid Files

pLenti-Cas-Guide

Lentiviral vector for co-expressing a guide RNA (gRNA) together with Cas9, for genome editing using the S. pyogenes CRISPR/Cas9 system.

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pLenti-Cas-Guide Sequence and MappLenti-Cas-Guide.dna
Map and Sequence File   
Sequence Author:  OriGene
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 FLAG SV40 NLS SV40 NLS CMV enhancer CF3 Primer (2745 .. 2766) BamHI (2951) BsmBI (3001) gRNA scaffold polIII terminator CMV enhancer T7 promoter pLenti-Cas-Guide 11,015 bp
BamHI  (2951)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
BsmBI  (3001)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
CF3 Primer
22-mer  /  45% GC
1 binding site
2745 .. 2766  =  22 annealed bases
Tm  =  56°C
Cas9
3814 .. 7917  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
3814 .. 7917  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
CmR
10,223 .. 10,882  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
10,223 .. 10,882  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
8199 .. 8787  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
8199 .. 8787  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
9558 .. 10,146  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
9558 .. 10,146  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
3128 .. 3507  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
3128 .. 3507  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
2707 .. 2947  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2707 .. 2947  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
CMV promoter
3508 .. 3711  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
3508 .. 3711  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
9316 .. 9496  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
9316 .. 9496  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
10,883 .. 10,987  =  105 bp
AmpR promoter
10,883 .. 10,987  =  105 bp
gRNA scaffold
3001 .. 3076  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
3001 .. 3076  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
Myc
7987 .. 8016  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
7987 .. 8016  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
8035 .. 8058  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
FLAG
8035 .. 8058  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
SV40 NLS
7918 .. 7938  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
7918 .. 7938  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
7945 .. 7965  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
7945 .. 7965  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
T7 promoter
3737 .. 3755  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3737 .. 3755  =  19 bp
promoter for bacteriophage T7 RNA polymerase
polIII terminator
3077 .. 3082  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3077 .. 3082  =  6 bp
RNA polymerase III transcription terminator
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