pCas9D10A_GFP

Plasmid for co-expression in mammalian cells of enhanced GFP (EGFP) and human codon-optimized Cas9 with the D10A nickase mutation.

Sequence Author: Musunuru Lab / Addgene #44720

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pCas9D10A_GFP 9271 bp
Cas9(D10A)
1745 .. 5848  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 1:  
   1745 .. 1771  =  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)
1745 .. 5848  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 2:  
   1772 .. 1774  =  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)
1745 .. 5848  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 3:  
   1775 .. 5848  =  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)
1745 .. 5848  =  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
5861 .. 5881  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5861 .. 5881  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
T2A
5897 .. 5950  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
T2A
5897 .. 5950  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
EGFP
5951 .. 6670  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   5951 .. 5953  =  3 bp
   1 amino acid  =  149.2 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
5951 .. 6670  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   5954 .. 5956  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
5951 .. 6670  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   5957 .. 6670  =  714 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
5951 .. 6670  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: enhanced GFP
mammalian codon-optimized
chimeric intron
666 .. 1678  =  1013 bp
chimera between introns from chicken β-actin and rabbit β-globin
chimeric intron
666 .. 1678  =  1013 bp
chimera between introns from chicken β-actin and rabbit β-globin
AmpR
8044 .. 8904  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8044 .. 8835  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8044 .. 8904  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8836 .. 8904  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8044 .. 8904  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7285 .. 7873  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7285 .. 7873  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
8 .. 387  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
8 .. 387  =  380 bp
human cytomegalovirus immediate early enhancer
chicken β-actin promoter
390 .. 665  =  276 bp
chicken β-actin promoter
390 .. 665  =  276 bp
SV40 poly(A) signal
6713 .. 6834  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6713 .. 6834  =  122 bp
SV40 polyadenylation signal
AmpR promoter
8905 .. 9009  =  105 bp
AmpR promoter
8905 .. 9009  =  105 bp
ORF:  259 .. 738  =  480 bp
ORF:  159 amino acids  =  17.1 kDa
ORF:  1490 .. 6670  =  5181 bp
ORF:  1726 amino acids  =  197.9 kDa
ORF:  8174 .. 8440  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1650 .. 1874  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  806 .. 1351  =  546 bp
ORF:  181 amino acids  =  19.3 kDa
ORF:  1358 .. 1657  =  300 bp
ORF:  99 amino acids  =  10.5 kDa
ORF:  1673 .. 2224  =  552 bp
ORF:  183 amino acids  =  20.5 kDa
ORF:  3149 .. 3667  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  3731 .. 4264  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
ORF:  5258 .. 5791  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  8044 .. 8904  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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