pCAG-T3-hCAS-pA

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

Sequence Author: Fujii

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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
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
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
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
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
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
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
2 segments
   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
2 segments
   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
T3 promoter
5555 .. 5573  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
5555 .. 5573  =  19 bp
promoter for bacteriophage T3 RNA polymerase
SV40 ori
160 .. 295  =  136 bp
SV40 origin of replication
SV40 ori
160 .. 295  =  136 bp
SV40 origin of replication
ORF:  7420 .. 8280  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  944 .. 1369  =  426 bp
ORF:  141 amino acids  =  16.8 kDa
ORF:  1373 .. 1906  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  2900 .. 3433  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
ORF:  3497 .. 4015  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  4940 .. 5635  =  696 bp
ORF:  231 amino acids  =  25.8 kDa
ORF:  5651 .. 5950  =  300 bp
ORF:  99 amino acids  =  10.5 kDa
ORF:  5957 .. 6565  =  609 bp
ORF:  202 amino acids  =  21.2 kDa
ORF:  6669 .. 6995  =  327 bp
ORF:  108 amino acids  =  11.4 kDa
ORF:  7884 .. 8150  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1280 .. 5497  =  4218 bp
ORF:  1405 amino acids  =  162.6 kDa
ORF:  5558 .. 5818  =  261 bp
ORF:  86 amino acids  =  9.4 kDa
ORF:  6575 .. 6805  =  231 bp
ORF:  76 amino acids  =  8.2 kDa
ORF:  5290 .. 5658  =  369 bp
ORF:  122 amino acids  =  13.3 kDa
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