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Plasmid Files

pX602

Zhang lab pX602-AAV-TBG::NLS-SaCas9-NLS-HA-OLLAS-bGHpA;U6::BsaI-sgRNA vector, for expressing S. aureus Cas9 plus a single guide RNA. 

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pX602 Sequence and MappX602.dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #61593
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 U6 promoter BsaI (4298) BsaI (4274) bGH poly(A) signal HA ATG pX602 7293 bp
BsaI  (4298)
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  (4274)
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
672 .. 3827  =  3156 bp
1052 amino acids  =  123.8 kDa
Cas9 (Csn1) endonuclease from the Staphylococcus
aureus
Type II CRISPR/Cas system
SaCas9
672 .. 3827  =  3156 bp
1052 amino acids  =  123.8 kDa
Cas9 (Csn1) endonuclease from the Staphylococcus
aureus
Type II CRISPR/Cas system
AmpR
5613 .. 6473  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5613 .. 5681  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5613 .. 6473  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5682 .. 6473  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5613 .. 6473  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
6644 .. 7232  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6644 .. 7232  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4771 .. 5226  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4771 .. 5226  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
TBG promoter
187 .. 596  =  410 bp
promoter from the human thyroxine-binding
globulin gene (Yan et al., 2012)
TBG promoter
187 .. 596  =  410 bp
promoter from the human thyroxine-binding
globulin gene (Yan et al., 2012)
U6 promoter
4307 .. 4547  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
4307 .. 4547  =  241 bp
RNA polymerase III promoter for human U6 snRNA
bGH poly(A) signal
3984 .. 4191  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
3984 .. 4191  =  208 bp
bovine growth hormone polyadenylation signal
AAV2 ITR
4556 .. 4696  =  141 bp
inverted terminal repeat of adeno-associated virus
serotype 2
AAV2 ITR
4556 .. 4696  =  141 bp
inverted terminal repeat of adeno-associated virus
serotype 2
AmpR promoter
5508 .. 5612  =  105 bp
AmpR promoter
5508 .. 5612  =  105 bp
Sa gRNA scaffold
4203 .. 4278  =  76 bp
guide RNA scaffold for the Staphylococcus aureus
CRISPR/Cas9 system
Sa gRNA scaffold
4203 .. 4278  =  76 bp
guide RNA scaffold for the Staphylococcus aureus
CRISPR/Cas9 system
nucleoplasmin NLS
3828 .. 3875  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
3828 .. 3875  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
OLLAS tag
3909 .. 3950  =  42 bp
14 amino acids  =  1.5 kDa
Product: Escherichia coli OmpF Linker and mouse
Langerin fusion sequence tag (Park et al., 2008)
OLLAS tag
3909 .. 3950  =  42 bp
14 amino acids  =  1.5 kDa
Product: Escherichia coli OmpF Linker and mouse
Langerin fusion sequence tag (Park et al., 2008)
HA
3882 .. 3908  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
HA
3882 .. 3908  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
SV40 NLS
627 .. 647  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
627 .. 647  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
ATG
621 .. 623  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
621 .. 623  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
K135R
1071 .. 1073  =  3 bp
Lys to Arg mutation
K135R
1071 .. 1073  =  3 bp
Lys to Arg mutation
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