pST1374-N-NLS-flag-linker-Cas9-D10A

Plasmid for mammalian cell expression of Cas9 with the D10A nickase mutation and with an N-terminal nuclear localization signal followed by a FLAG® tag.

Sequence Author: Huang Lab / Addgene #51130

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CMV enhancer ATG SV40 NLS pST1374-N-NLS-flag-linker-Cas9-D10A 9317 bp
Cas9(D10A)
2212 .. 6315  =  4104 bp
1367 amino acids  =  158.3 kDa
3 segments
   Segment 3:  
   2212 .. 6288  =  4077 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)
2212 .. 6315  =  4104 bp
1367 amino acids  =  158.3 kDa
3 segments
   Segment 2:  
   6289 .. 6291  =  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)
2212 .. 6315  =  4104 bp
1367 amino acids  =  158.3 kDa
3 segments
   Segment 1:  
   6292 .. 6315  =  24 bp
   8 amino acids  =  923.1 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)
2212 .. 6315  =  4104 bp
1367 amino acids  =  158.3 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
FLAG
6418 .. 6441  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
6418 .. 6441  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
SV40 NLS
6451 .. 6471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6451 .. 6471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
ATG
6475 .. 6477  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
6475 .. 6477  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
AmpR
7517 .. 8377  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7517 .. 7585  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7517 .. 8377  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7586 .. 8377  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7517 .. 8377  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
8548 .. 9136  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
8548 .. 9136  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1404 .. 1832  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1404 .. 1832  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
549 .. 947  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
549 .. 947  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
6767 .. 7146  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
6767 .. 7146  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
1061 .. 1390  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1061 .. 1390  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
1878 .. 2102  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1878 .. 2102  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
6563 .. 6766  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
6563 .. 6766  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
270 .. 391  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
270 .. 391  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7412 .. 7516  =  105 bp
AmpR promoter
7412 .. 7516  =  105 bp
EM7 promoter
966 .. 1013  =  48 bp
synthetic bacterial promoter
EM7 promoter
966 .. 1013  =  48 bp
synthetic bacterial promoter
T7 promoter
6500 .. 6518  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
6500 .. 6518  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
1075 .. 1210  =  136 bp
SV40 origin of replication
SV40 ori
1075 .. 1210  =  136 bp
SV40 origin of replication
ORF:  2533 .. 3003  =  471 bp
ORF:  156 amino acids  =  17.7 kDa
ORF:  3370 .. 3705  =  336 bp
ORF:  111 amino acids  =  12.7 kDa
ORF:  3799 .. 4131  =  333 bp
ORF:  110 amino acids  =  12.7 kDa
ORF:  5332 .. 5664  =  333 bp
ORF:  110 amino acids  =  12.0 kDa
ORF:  5839 .. 6195  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  6196 .. 6519  =  324 bp
ORF:  107 amino acids  =  11.8 kDa
ORF:  2240 .. 2527  =  288 bp
ORF:  95 amino acids  =  10.7 kDa
ORF:  7517 .. 8377  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  645 .. 1001  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  549 .. 947  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  1615 .. 1881  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  2212 .. 6477  =  4266 bp
ORF:  1421 amino acids  =  164.4 kDa
ORF:  7981 .. 8247  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pST1374-N-NLS-flag-linker-Cas9-D10A.dna file

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