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Plasmid Files

pX601

Zhang lab pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA vector, for expressing S. aureus Cas9 plus a single guide RNA. 

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pX601 Sequence and MappX601.dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #61591
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 BsaI (4619) BsaI (4595) bGH poly(A) signal ATG pX601 7446 bp
BsaI  (4619)
2 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BsaI  (4595)
2 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
SaCas9
813 .. 3968  =  3156 bp
1052 amino acids  =  123.8 kDa
Product: Cas9 (Csn1) endonuclease from the
Staphylococcus aureus Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SaCas9
813 .. 3968  =  3156 bp
1052 amino acids  =  123.8 kDa
Product: Cas9 (Csn1) endonuclease from the
Staphylococcus aureus Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
AmpR
5766 .. 6626  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5766 .. 5834  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5766 .. 6626  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5835 .. 6626  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5766 .. 6626  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
6797 .. 7385  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6797 .. 7385  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4924 .. 5379  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4924 .. 5379  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
154 .. 533  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
154 .. 533  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
4351 .. 4591  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
4351 .. 4591  =  241 bp
RNA polymerase III promoter for human U6 snRNA
bGH poly(A) signal
4137 .. 4344  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
4137 .. 4344  =  208 bp
bovine growth hormone polyadenylation signal
CMV promoter
534 .. 737  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
534 .. 737  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
AAV2 ITR
4709 .. 4849  =  141 bp
inverted terminal repeat of adeno-associated virus
serotype 2
AAV2 ITR
4709 .. 4849  =  141 bp
inverted terminal repeat of adeno-associated virus
serotype 2
AmpR promoter
5661 .. 5765  =  105 bp
AmpR promoter
5661 .. 5765  =  105 bp
3xHA
4023 .. 4103  =  81 bp
27 amino acids  =  3.3 kDa
Product: three tandem HA epitope tags
3xHA
4023 .. 4103  =  81 bp
27 amino acids  =  3.3 kDa
Product: three tandem HA epitope tags
Sa gRNA scaffold
4620 .. 4695  =  76 bp
guide RNA scaffold for the Staphylococcus aureus
CRISPR/Cas9 system
Sa gRNA scaffold
4620 .. 4695  =  76 bp
guide RNA scaffold for the Staphylococcus aureus
CRISPR/Cas9 system
nucleoplasmin NLS
3969 .. 4016  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
3969 .. 4016  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
SV40 NLS
768 .. 788  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
768 .. 788  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
ATG
762 .. 764  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
762 .. 764  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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