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Plasmid Files

pT7-Cas9-Nickase

In vitro transcription vector with the T7 promoter, for producing myc- and FLAG®-tagged S. pyogenes Cas9 with the D10A nickase mutation.

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pT7-Cas9-Nickase.dna
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Sequence Author:  OriGene
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PmeI (5277) pT7-Cas9-Nickase 9036 bp
PmeI  (5277)
1 site
G T T T A A A C C A A A T T T G
Cas9(D10A)
1029 .. 5132  =  4104 bp
1368 amino acids  =  158.4 kDa
   Segment 1:  
   1029 .. 1055  =  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)
1029 .. 5132  =  4104 bp
1368 amino acids  =  158.4 kDa
   Segment 2:  
   1056 .. 1058  =  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)
1029 .. 5132  =  4104 bp
1368 amino acids  =  158.4 kDa
   Segment 3:  
   1059 .. 5132  =  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)
1029 .. 5132  =  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
NeoR/KanR
7287 .. 8081  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
7287 .. 8081  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
hGH poly(A) signal
5320 .. 5942  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
5320 .. 5942  =  623 bp
human growth hormone polyadenylation signal
ori
6091 .. 6679  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6091 .. 6679  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
8606 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
8606 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
8116 .. 8473  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
8116 .. 8473  =  358 bp
SV40 enhancer and early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
AmpR promoter
8475 .. 8579  =  105 bp
AmpR promoter
8475 .. 8579  =  105 bp
HSV TK poly(A) signal
7008 .. 7055  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
7008 .. 7055  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
Myc
5202 .. 5231  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
5202 .. 5231  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
5250 .. 5273  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
5250 .. 5273  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
SV40 NLS
5133 .. 5153  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5133 .. 5153  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5160 .. 5180  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5160 .. 5180  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  882 .. 5279  =  4398 bp
ORF:  1465 amino acids  =  169.7 kDa
ORF:  6828 .. 7061  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  7236 .. 7772  =  537 bp
ORF:  178 amino acids  =  19.9 kDa
ORF:  6817 .. 7266  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  1287 .. 1529  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
ORF:  3081 .. 3425  =  345 bp
ORF:  114 amino acids  =  13.5 kDa
ORF:  3579 .. 3983  =  405 bp
ORF:  134 amino acids  =  15.6 kDa
ORF:  4452 .. 5075  =  624 bp
ORF:  207 amino acids  =  24.3 kDa
ORF:  7287 .. 8081  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5684 .. 5908  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  7523 .. 7909  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
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