pST1374-N-NLS-flag-linker-Cas9-H840A

Plasmid for mammalian cell expression of Cas9 with the H840A nickase mutation and with an N-terminal nuclear localization signal followed by a FLAG® tag.

Sequence Author: Huang Lab / Addgene #51129

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CMV enhancer ATG SV40 NLS pST1374-N-NLS-flag-linker-Cas9-H840A 9318 bp
ATG
904 .. 906  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
904 .. 906  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
910 .. 930  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
910 .. 930  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
FLAG
940 .. 963  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
940 .. 963  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
Cas9(H840A)
1066 .. 5169  =  4104 bp
1367 amino acids  =  158.2 kDa
3 segments
   Segment 1:  
   1066 .. 3579  =  2514 bp
   838 amino acids  =  97.3 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 H840A mutation in the HNH catalytic domain
Cas9(H840A)
1066 .. 5169  =  4104 bp
1367 amino acids  =  158.2 kDa
3 segments
   Segment 2:  
   3580 .. 3582  =  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 H840A mutation in the HNH catalytic domain
Cas9(H840A)
1066 .. 5169  =  4104 bp
1367 amino acids  =  158.2 kDa
3 segments
   Segment 3:  
   3583 .. 5169  =  1587 bp
   528 amino acids  =  60.9 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 H840A mutation in the HNH catalytic domain
Cas9(H840A)
1066 .. 5169  =  4104 bp
1367 amino acids  =  158.2 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 H840A mutation in the HNH catalytic domain
AmpR
8322 .. 9182  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8322 .. 9113  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8322 .. 9182  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   9114 .. 9182  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8322 .. 9182  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7563 .. 8151  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7563 .. 8151  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5549 .. 5977  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5549 .. 5977  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
6435 .. 6833  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
6435 .. 6833  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
5991 .. 6321  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
5991 .. 6321  =  331 bp
SV40 enhancer and early promoter
bGH poly(A) signal
5279 .. 5503  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5279 .. 5503  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
6991 .. 7112  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6991 .. 7112  =  122 bp
SV40 polyadenylation signal
AmpR promoter
9183 .. 9287  =  105 bp
AmpR promoter
9183 .. 9287  =  105 bp
EM7 promoter
6369 .. 6416  =  48 bp
synthetic bacterial promoter
EM7 promoter
6369 .. 6416  =  48 bp
synthetic bacterial promoter
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
6172 .. 6307  =  136 bp
SV40 origin of replication
SV40 ori
6172 .. 6307  =  136 bp
SV40 origin of replication
ORF:  904 .. 5169  =  4266 bp
ORF:  1421 amino acids  =  164.4 kDa
ORF:  5500 .. 5766  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  8452 .. 8718  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6435 .. 6833  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  862 .. 1185  =  324 bp
ORF:  107 amino acids  =  11.9 kDa
ORF:  1186 .. 1542  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  1717 .. 2049  =  333 bp
ORF:  110 amino acids  =  12.0 kDa
ORF:  3676 .. 4011  =  336 bp
ORF:  111 amino acids  =  12.7 kDa
ORF:  4378 .. 4848  =  471 bp
ORF:  156 amino acids  =  17.7 kDa
ORF:  4854 .. 5141  =  288 bp
ORF:  95 amino acids  =  10.7 kDa
ORF:  6381 .. 6737  =  357 bp
ORF:  118 amino acids  =  12.9 kDa
ORF:  8322 .. 9182  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pST1374-N-NLS-flag-linker-Cas9-H840A.dna file

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