Resources
Plasmid Files

pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s)

Gene editing plasmid for expressing Cas9, tracrRNA, and a crRNA using the Neisseria meningitidis CRISPR/Cas9 system.

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

pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) Sequence and MappSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s).dna
Map and Sequence File   
Sequence Author:  Thomson & Sontheimer Labs / Addgene #47868
Download Free Trial Get SnapGene Viewer

 BsmBI (11) NLS BsmBI (35) ATG pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) 9258 bp
BsmBI  (11)
2 sites
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.
BsmBI  (35)
2 sites
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.
NmCas9
2205 .. 5447  =  3243 bp
1081 amino acids  =  124.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Neisseria meningitidis Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
NmCas9
2205 .. 5447  =  3243 bp
1081 amino acids  =  124.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Neisseria meningitidis Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
EF-1α promoter
993 .. 2166  =  1174 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
993 .. 2166  =  1174 bp
strong constitutive promoter for human elongation
factor EF-1α
mini-oriP
5942 .. 7055  =  1114 bp
internally truncated variant of the Epstein-Barr virus
oriP replication origin (Tanaka et al., 1999)
mini-oriP
5942 .. 7055  =  1114 bp
internally truncated variant of the Epstein-Barr virus
oriP replication origin (Tanaka et al., 1999)
AmpR
8262 .. 9122  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   8262 .. 9053  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8262 .. 9122  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   9054 .. 9122  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8262 .. 9122  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
7503 .. 8091  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7503 .. 8091  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
U6 promoter
44 .. 284  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
44 .. 284  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
519 .. 759  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
519 .. 759  =  241 bp
RNA polymerase III promoter for human U6 snRNA
bGH poly(A) signal
5529 .. 5753  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5529 .. 5753  =  225 bp
bovine growth hormone polyadenylation signal
AmpR promoter
9123 .. 9227  =  105 bp
AmpR promoter
9123 .. 9227  =  105 bp
Nm tracrRNA
419 .. 511  =  93 bp
trans-activating CRISPR RNA for the Neisseria
meningitidis
CRISPR/Cas9 system (Hou et al., 2013)
Nm tracrRNA
419 .. 511  =  93 bp
trans-activating CRISPR RNA for the Neisseria
meningitidis
CRISPR/Cas9 system (Hou et al., 2013)
HA
5448 .. 5474  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
HA
5448 .. 5474  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
NLS
5481 .. 5507  =  27 bp
9 amino acids  =  998.3 Da
Product: nuclear localization signal (Makkerh et al.,
1996)
NLS
5481 .. 5507  =  27 bp
9 amino acids  =  998.3 Da
Product: nuclear localization signal (Makkerh et al.,
1996)
SV40 NLS
2184 .. 2204  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
2184 .. 2204  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
ATG
2178 .. 2180  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2178 .. 2180  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
EF-1α intron A
1224 .. 2157  =  934 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
1224 .. 2157  =  934 bp
intron upstream of the start codon of human EF-1α
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 © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter