Resources
Plasmid Files

pCas-Guide-Nickase

Mammalian vector for co-expressing a guide RNA (gRNA) together with the D10A nickase mutant of Cas9, for genome editing using the CRISPR/Cas9 system.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pCas-Guide-Nickase.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
Download Free Trial Get SnapGene Viewer


CF3 Primer (128 .. 149) BamHI (334) BsmBI (384) pCas-Guide-Nickase 8033 bp
BamHI  (334)
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  (384)
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
128 .. 149  =  22 annealed bases
Tm  =  56°C
primer for sequencing the inserted gRNA
Cas9(D10A)
1197 .. 5300  =  4104 bp
1368 amino acids  =  158.4 kDa
   Segment 1:  
   1197 .. 1223  =  27 bp
   9 amino acids  =  1.1 kDa
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
Cas9(D10A)
1197 .. 5300  =  4104 bp
1368 amino acids  =  158.4 kDa
   Segment 2:  
   1224 .. 1226  =  3 bp
   1 amino acid  =  89.1 Da
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
Cas9(D10A)
1197 .. 5300  =  4104 bp
1368 amino acids  =  158.4 kDa
   Segment 3:  
   1227 .. 5300  =  4074 bp
   1358 amino acids  =  157.3 kDa
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
Cas9(D10A)
1197 .. 5300  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
AmpR
6994 .. 7854  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6994 .. 7785  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6994 .. 7854  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7786 .. 7854  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6994 .. 7854  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
hGH poly(A) signal
5488 .. 6110  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
5488 .. 6110  =  623 bp
human growth hormone polyadenylation signal
ori
6231 .. 6819  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6231 .. 6819  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
511 .. 890  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
511 .. 890  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
90 .. 330  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
90 .. 330  =  241 bp
RNA polymerase III promoter for human U6 snRNA
CMV promoter
891 .. 1094  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
891 .. 1094  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
gRNA scaffold
384 .. 459  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
384 .. 459  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
Myc
5370 .. 5399  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
5370 .. 5399  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
5418 .. 5441  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
5418 .. 5441  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
SV40 NLS
5301 .. 5321  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5301 .. 5321  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5328 .. 5348  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5328 .. 5348  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
T7 promoter
1120 .. 1138  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1120 .. 1138  =  19 bp
promoter for bacteriophage T7 RNA polymerase
polIII terminator
460 .. 465  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
460 .. 465  =  6 bp
RNA polymerase III transcription terminator
ORF:  6200 .. 6556  =  357 bp
ORF:  118 amino acids  =  13.2 kDa
ORF:  7124 .. 7390  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1050 .. 5447  =  4398 bp
ORF:  1465 amino acids  =  169.8 kDa
ORF:  1455 .. 1697  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
ORF:  3249 .. 3593  =  345 bp
ORF:  114 amino acids  =  13.5 kDa
ORF:  3747 .. 4151  =  405 bp
ORF:  134 amino acids  =  15.6 kDa
ORF:  4620 .. 5243  =  624 bp
ORF:  207 amino acids  =  24.3 kDa
ORF:  5852 .. 6076  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  6994 .. 7854  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2018 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter