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Plasmid Files

pT3TS-nCas9n

Plasmid for T3 promoter-driven expression of zebrafish codon-optimized S. pyogenes Cas9.

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pT3TS-nCas9n Sequence and MappT3TS-nCas9n.dna
Map and Sequence File   
Sequence Author:  Chen Lab / Addgene #46757
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 T3 promoter XbaI (4500) ATG pT3TS-nCas9n 7330 bp
XbaI  (4500)
1 site
T C T A G A A G A T C T
Cas9
147 .. 4247  =  4101 bp
1367 amino acids  =  158.2 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
147 .. 4247  =  4101 bp
1367 amino acids  =  158.2 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
AmpR
5289 .. 6149  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5289 .. 5357  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5289 .. 6149  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5358 .. 6149  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5289 .. 6149  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
6320 .. 6908  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6320 .. 6908  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4702 .. 5157  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4702 .. 5157  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
Xenopus globin 3'-UTR
4308 .. 4433  =  126 bp
3'-UTR of the major beta-globin gene of Xenopus
laevis
Xenopus globin 3'-UTR
4308 .. 4433  =  126 bp
3'-UTR of the major beta-globin gene of Xenopus
laevis
AmpR promoter
5184 .. 5288  =  105 bp
AmpR promoter
5184 .. 5288  =  105 bp
Xenopus globin 5'-UTR
44 .. 86  =  43 bp
translational enhancer from the 5'-UTR of the major
beta-globin gene of Xenopus laevis
Xenopus globin 5'-UTR
44 .. 86  =  43 bp
translational enhancer from the 5'-UTR of the major
beta-globin gene of Xenopus laevis
SV40 NLS
120 .. 140  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
120 .. 140  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
4263 .. 4283  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
4263 .. 4283  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
T3 promoter
1 .. 19  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
1 .. 19  =  19 bp
promoter for bacteriophage T3 RNA polymerase
ATG
108 .. 110  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
108 .. 110  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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