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Plasmid Files

PX552

Zhang lab AAV vector, also known as AAV-SpGuide or pAAV SpCas9 acceptor, for sgRNA expression with a nuclear EGFP marker. Use together with PX551.

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PX552 Sequence and MapPX552.dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #60958
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 SapI (401) SapI (422) pol III terminator PX552 5303 bp
SapI  (401)
2 sites
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
SapI  (422)
2 sites
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
AmpR
3623 .. 4483  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3623 .. 3691  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3623 .. 4483  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3692 .. 4483  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3623 .. 4483  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
EGFP
1017 .. 1733  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 1:  
   1017 .. 1019  =  3 bp
   1 amino acid  =  149.2 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
1017 .. 1733  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 2:  1a  
   1020 .. 1022  =  3 bp
   1 amino acid  =  117.2 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
1017 .. 1733  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 3:  
   1023 .. 1733  =  711 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
1017 .. 1733  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
Product: enhanced GFP
mammalian codon-optimized
ori
4654 .. 5242  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4654 .. 5242  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hGH poly(A) signal
2050 .. 2526  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
2050 .. 2526  =  477 bp
human growth hormone polyadenylation signal
f1 ori
2781 .. 3236  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2781 .. 3236  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
hSyn promoter
532 .. 979  =  448 bp
human synapsin I promoter; confers neuron-specific
expression (Kügler et al., 2003)
hSyn promoter
532 .. 979  =  448 bp
human synapsin I promoter; confers neuron-specific
expression (Kügler et al., 2003)
KASH
1752 .. 2000  =  249 bp
82 amino acids  =  9.2 kDa
Product: transmembrane KASH domain of nesprin-2
confers localization to the nuclear envelope
KASH
1752 .. 2000  =  249 bp
82 amino acids  =  9.2 kDa
Product: transmembrane KASH domain of nesprin-2
confers localization to the nuclear envelope
U6 promoter
156 .. 396  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
156 .. 396  =  241 bp
RNA polymerase III promoter for human U6 snRNA
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
AAV2 ITR
2566 .. 2706  =  141 bp
inverted terminal repeat of adeno-associated virus
serotype 2
AAV2 ITR
2566 .. 2706  =  141 bp
inverted terminal repeat of adeno-associated virus
serotype 2
AmpR promoter
3518 .. 3622  =  105 bp
AmpR promoter
3518 .. 3622  =  105 bp
gRNA scaffold
423 .. 498  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
423 .. 498  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
pol III terminator
499 .. 504  =  6 bp
RNA polymerase III transcription terminator
pol III terminator
499 .. 504  =  6 bp
RNA polymerase III transcription terminator
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