SP-Cas9

Bacterial plasmid for regulated expression of bacterial codon-optimized S. pyogenes Cas9 with a C-terminal nuclear localization signal and 6xHis tag.

Sequence Author: Geijsen Lab / Addgene #62731

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SV40 NLS RBS 6xHis T7 promoter SP-Cas9 9822 bp
Cas9
5317 .. 9420  =  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
5317 .. 9420  =  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
SV40 NLS
9445 .. 9465  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
9445 .. 9465  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
6xHis
9478 .. 9495  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
9478 .. 9495  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lacI
3761 .. 4843  =  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
3761 .. 4843  =  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
209 .. 1069  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   209 .. 277  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
209 .. 1069  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   278 .. 1069  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
209 .. 1069  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1240 .. 1828  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1240 .. 1828  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
2258 .. 2449  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
2258 .. 2449  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
104 .. 208  =  105 bp
AmpR promoter
104 .. 208  =  105 bp
lacI promoter
4844 .. 4921  =  78 bp
lacI promoter
4844 .. 4921  =  78 bp
T7 terminator
9563 .. 9610  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
9563 .. 9610  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
5249 .. 5273  =  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
5249 .. 5273  =  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
5288 .. 5310  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
5288 .. 5310  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
5230 .. 5248  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5230 .. 5248  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  3283 .. 3978  =  696 bp
ORF:  231 amino acids  =  25.4 kDa
ORF:  4882 .. 5145  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  5317 .. 9498  =  4182 bp
ORF:  1393 amino acids  =  161.1 kDa
ORF:  209 .. 1069  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2481 .. 2849  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  5019 .. 5258  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  673 .. 939  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3088 .. 3705  =  618 bp
ORF:  205 amino acids  =  21.8 kDa
ORF:  6268 .. 7101  =  834 bp
ORF:  277 amino acids  =  33.1 kDa
ORF:  7504 .. 8106  =  603 bp
ORF:  200 amino acids  =  23.8 kDa
ORF:  8434 .. 9102  =  669 bp
ORF:  222 amino acids  =  26.9 kDa
ORF:  3741 .. 4004  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4593 .. 5093  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  8430 .. 8822  =  393 bp
ORF:  130 amino acids  =  14.5 kDa
ORF:  2258 .. 2482  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  2846 .. 3184  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  3761 .. 4720  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  6443 .. 6706  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
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