pSpCas9 (PX165)

Zhang lab vector for expressing human codon-optimized Cas9. sgRNA sequences can be co-transfected as PCR products.

Sequence Author: Zhang Lab / Addgene #48137

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nucleoplasmin NLS hybrid intron ATG SV40 NLS pSpCas9 (PX165) 8226 bp
ATG
971 .. 973  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
971 .. 973  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
974 .. 1039  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
974 .. 1039  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
1046 .. 1066  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1046 .. 1066  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
1091 .. 5191  =  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
1091 .. 5191  =  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
nucleoplasmin NLS
5192 .. 5239  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
5192 .. 5239  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
AmpR
6546 .. 7406  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6546 .. 6614  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6546 .. 7406  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6615 .. 7406  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6546 .. 7406  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7577 .. 8165  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7577 .. 8165  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5704 .. 6159  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5704 .. 6159  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
160 .. 445  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
CMV enhancer
160 .. 445  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
chicken β-actin promoter
447 .. 724  =  278 bp
chicken β-actin promoter
447 .. 724  =  278 bp
hybrid intron
725 .. 953  =  229 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
hybrid intron
725 .. 953  =  229 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
bGH poly(A) signal
5273 .. 5480  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5273 .. 5480  =  208 bp
bovine growth hormone polyadenylation signal
AAV2 ITR
5489 .. 5629  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
5489 .. 5629  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
1 .. 141  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
1 .. 141  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
6441 .. 6545  =  105 bp
AmpR promoter
6441 .. 6545  =  105 bp
ORF:  355 .. 702  =  348 bp
ORF:  115 amino acids  =  12.1 kDa
ORF:  5503 .. 5727  =  225 bp
ORF:  74 amino acids  =  8.3 kDa
ORF:  545 .. 856  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  971 .. 5242  =  4272 bp
ORF:  1423 amino acids  =  164.6 kDa
ORF:  909 .. 1148  =  240 bp
ORF:  79 amino acids  =  9.1 kDa
ORF:  6546 .. 7406  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2437 .. 2730  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  3001 .. 3450  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  5403 .. 5684  =  282 bp
ORF:  93 amino acids  =  10.4 kDa
ORF:  950 .. 3607  =  2658 bp
ORF:  885 amino acids  =  93.1 kDa
ORF:  7010 .. 7276  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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