pET-NLS-Cas9-6xHis

Bacterial vector for regulated expression of S. pyogenes Cas9 with an N-terminal nuclear localization signal and C-terminal 6xHis tag.

Sequence Author: Liu Lab / Addgene #62934

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T7 promoter RBS ATG SV40 NLS pET-NLS-Cas9-6xHis 9289 bp
ATG
71 .. 73  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
71 .. 73  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
74 .. 94  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
74 .. 94  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
95 .. 4198  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
95 .. 4198  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
6xHis
4199 .. 4216  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4199 .. 4216  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lacI
7800 .. 8882  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
7800 .. 8882  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
AmpR
4985 .. 5845  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4985 .. 5776  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4985 .. 5845  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5777 .. 5845  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4985 .. 5845  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
6019 .. 6607  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6019 .. 6607  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4415 .. 4870  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4415 .. 4870  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
7037 .. 7228  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
7037 .. 7228  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
5846 .. 5938  =  93 bp
AmpR promoter
5846 .. 5938  =  93 bp
lacI promoter
8883 .. 8960  =  78 bp
lacI promoter
8883 .. 8960  =  78 bp
T7 terminator
4331 .. 4378  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
4331 .. 4378  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
3 .. 27  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
3 .. 27  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
RBS
42 .. 64  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
42 .. 64  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
9273 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
9273 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  9058 .. 20  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
ORF:  71 .. 4219  =  4149 bp
ORF:  1382 amino acids  =  160.3 kDa
ORF:  7766 .. 8017  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  8921 .. 9184  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  5115 .. 5381  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7260 .. 7628  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  4985 .. 5845  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  7037 .. 7261  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  7780 .. 8043  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  8632 .. 9132  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  7800 .. 8759  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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Download pET-NLS-Cas9-6xHis.dna file

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