lentiCas9n(D10A)-Blast

Zhang lab lentiviral vector with a blasticidin resistance marker, for expressing Cas9 with the D10A nickase mutation.

Sequence Author: Zhang Lab / Addgene #63593

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BleoR EM7 promoter 5' LTR (truncated) FLAG CMV enhancer lentiCas9n(D10A)-Blast 12,860 bp
Cas9(D10A)
2887 .. 6990  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 1:  
   2887 .. 2913  =  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)
2887 .. 6990  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 2:  
   2914 .. 2916  =  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)
2887 .. 6990  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 3:  
   2917 .. 6990  =  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)
2887 .. 6990  =  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
nucleoplasmin NLS
6991 .. 7038  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
6991 .. 7038  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
FLAG
7039 .. 7062  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
7039 .. 7062  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
P2A
7072 .. 7128  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
P2A
7072 .. 7128  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
BSD
7129 .. 7527  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
7129 .. 7527  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
AmpR
11,867 .. 12,727  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   11,867 .. 12,658  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
11,867 .. 12,727  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   12,659 .. 12,727  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
11,867 .. 12,727  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
7552 .. 8140  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
7552 .. 8140  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
11,108 .. 11,696  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
11,108 .. 11,696  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
9147 .. 9575  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
9147 .. 9575  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
238 .. 617  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
238 .. 617  =  380 bp
human cytomegalovirus immediate early enhancer
BleoR
10,032 .. 10,406  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
10,032 .. 10,406  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
9589 .. 9918  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
9589 .. 9918  =  330 bp
SV40 enhancer and early promoter
RRE
1680 .. 1913  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1680 .. 1913  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
bGH poly(A) signal
8877 .. 9101  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
8877 .. 9101  =  225 bp
bovine growth hormone polyadenylation signal
EF-1α core promoter
2651 .. 2862  =  212 bp
core promoter for human elongation factor EF-1α
EF-1α core promoter
2651 .. 2862  =  212 bp
core promoter for human elongation factor EF-1α
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
5' LTR (truncated)
835 .. 1015  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
835 .. 1015  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
8665 .. 8845  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
8665 .. 8845  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1062 .. 1187  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
1062 .. 1187  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
10,536 .. 10,657  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
10,536 .. 10,657  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2440 .. 2557  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2440 .. 2557  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
12,728 .. 12,832  =  105 bp
AmpR promoter
12,728 .. 12,832  =  105 bp
EM7 promoter
9966 .. 10,013  =  48 bp
synthetic bacterial promoter
EM7 promoter
9966 .. 10,013  =  48 bp
synthetic bacterial promoter
SV40 ori
9769 .. 9904  =  136 bp
SV40 origin of replication
SV40 ori
9769 .. 9904  =  136 bp
SV40 origin of replication
ORF:  1558 .. 2445  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  2887 .. 7527  =  4641 bp
ORF:  1546 amino acids  =  176.9 kDa
ORF:  7642 .. 8691  =  1050 bp
ORF:  349 amino acids  =  39.4 kDa
ORF:  7106 .. 7348  =  243 bp
ORF:  80 amino acids  =  8.8 kDa
ORF:  7655 .. 8167  =  513 bp
ORF:  170 amino acids  =  18.0 kDa
ORF:  9098 .. 9364  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  10,032 .. 10,406  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  11,997 .. 12,263  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1473 .. 1853  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  4236 .. 4529  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  4800 .. 5249  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  10,025 .. 10,312  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  11,867 .. 12,727  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2566 .. 7431  =  4866 bp
ORF:  1621 amino acids  =  171.1 kDa
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