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.

Sequence Author: Thomson

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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-v2 is an improved version of BsmBI.
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.
BsmBI-v2 is an improved version of BsmBI.
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
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
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
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  =  998.2 Da
Product: nuclear localization signal (Makkerh et al., 1996)
NLS
5481 .. 5507  =  27 bp
9 amino acids  =  998.2 Da
Product: nuclear localization signal (Makkerh et al., 1996)
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
2 segments
   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
2 segments
   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)
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
ORF:  3949 .. 4203  =  255 bp
ORF:  84 amino acids  =  9.2 kDa
ORF:  8392 .. 8658  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5750 .. 6040  =  291 bp
ORF:  96 amino acids  =  10.6 kDa
ORF:  1533 .. 1856  =  324 bp
ORF:  107 amino acids  =  11.0 kDa
ORF:  2178 .. 5510  =  3333 bp
ORF:  1110 amino acids  =  127.6 kDa
ORF:  849 .. 1613  =  765 bp
ORF:  254 amino acids  =  27.9 kDa
ORF:  3924 .. 4502  =  579 bp
ORF:  192 amino acids  =  21.6 kDa
ORF:  8262 .. 9122  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1469 .. 1906  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  5519 .. 5881  =  363 bp
ORF:  120 amino acids  =  13.1 kDa
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