hCas9_D10A

Mammalian vector with a G418 resistance marker for expressing human codon-optimized Cas9 with the D10A nickase mutation.

Sequence Author: Church Lab / Addgene #41816

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hCas9_D10A 9553 bp
Cas9(D10A)
748 .. 4851  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 1:  
   748 .. 774  =  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)
748 .. 4851  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 2:  
   775 .. 777  =  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)
748 .. 4851  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 3:  
   778 .. 4851  =  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)
748 .. 4851  =  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
SV40 NLS
4864 .. 4884  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4864 .. 4884  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
AmpR
8372 .. 9232  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8372 .. 9163  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8372 .. 9232  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   9164 .. 9232  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8372 .. 9232  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
6071 .. 6865  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
6071 .. 6865  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
7613 .. 8201  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7613 .. 8201  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5233 .. 5661  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5233 .. 5661  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
50 .. 429  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
50 .. 429  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
5675 .. 6004  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
5675 .. 6004  =  330 bp
SV40 enhancer and early promoter
CMV promoter
430 .. 633  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
430 .. 633  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
7041 .. 7162  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7041 .. 7162  =  122 bp
SV40 polyadenylation signal
AmpR promoter
9233 .. 9337  =  105 bp
AmpR promoter
9233 .. 9337  =  105 bp
HSV TK poly(A) signal
4983 .. 5031  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
4983 .. 5031  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
SV40 ori
5855 .. 5990  =  136 bp
SV40 origin of replication
SV40 ori
5855 .. 5990  =  136 bp
SV40 origin of replication
ORF:  589 .. 4887  =  4299 bp
ORF:  1432 amino acids  =  166.3 kDa
ORF:  6071 .. 6865  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  6243 .. 6629  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  8502 .. 8768  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6380 .. 6634  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  8372 .. 9232  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  724 .. 1227  =  504 bp
ORF:  167 amino acids  =  18.5 kDa
ORF:  2152 .. 2670  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  2734 .. 3267  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
ORF:  4261 .. 4794  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  648 .. 890  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
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