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Plasmid Files

pCAG-T3-hCAS-pA

Plasmid for mammalian cell expression or in vitro transcription of human codon-optimized Cas9.

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pCAG-T3-hCAS-pA Sequence and MappCAG-T3-hCAS-pA.dna
Map and Sequence File   
Sequence Author:  Fujii & Naito Labs / Addgene #48625
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 ATG SV40 NLS SphI (1092) pCAG-T3-hCAS-pA 9272 bp
SphI  (1092)
1 site
G C A T G C C G T A C G
Cas9
1316 .. 5416  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
1316 .. 5416  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
chimeric intron
5630 .. 6646  =  1017 bp
chimera between introns from chicken β-actin and
rabbit β-globin
chimeric intron
5630 .. 6646  =  1017 bp
chimera between introns from chicken β-actin and
rabbit β-globin
AmpR
7420 .. 8280  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7420 .. 7488  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7420 .. 8280  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7489 .. 8280  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7420 .. 8280  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
8451 .. 9039  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
8451 .. 9039  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
6905 .. 7284  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
6905 .. 7284  =  380 bp
human cytomegalovirus immediate early enhancer
chicken β-actin promoter
6647 .. 6903  =  257 bp
contains a 21-bp deletion relative to the standard
chicken β-actin promoter
chicken β-actin promoter
6647 .. 6903  =  257 bp
contains a 21-bp deletion relative to the standard
chicken β-actin promoter
SV40 promoter
146 .. 341  =  196 bp
SV40 early promoter
SV40 promoter
146 .. 341  =  196 bp
SV40 early promoter
SV40 poly(A) signal
6 .. 140  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6 .. 140  =  135 bp
SV40 polyadenylation signal
AmpR promoter
7315 .. 7419  =  105 bp
AmpR promoter
7315 .. 7419  =  105 bp
poly(A)
1103 .. 1197  =  95 bp
enables microinjection of poly(A)-tailed mRNA after
in vitro transcription
poly(A)
1103 .. 1197  =  95 bp
enables microinjection of poly(A)-tailed mRNA after
in vitro transcription
3' UTR
1198 .. 1267  =  70 bp
portion of the 3' UTR from the human TBPL1 gene
3' UTR
1198 .. 1267  =  70 bp
portion of the 3' UTR from the human TBPL1 gene
β-globin poly(A) signal
875 .. 930  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
875 .. 930  =  56 bp
rabbit β-globin polyadenylation signal
FLAG
5471 .. 5494  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
FLAG
5471 .. 5494  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
SV40 NLS
1283 .. 1303  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
1283 .. 1303  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
5441 .. 5461  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
5441 .. 5461  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
T3 promoter
5555 .. 5573  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
5555 .. 5573  =  19 bp
promoter for bacteriophage T3 RNA polymerase
ATG
5495 .. 5497  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5495 .. 5497  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 ori
160 .. 295  =  136 bp
SV40 origin of replication
SV40 ori
160 .. 295  =  136 bp
SV40 origin of replication
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