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Plasmid Files

pRGE31

Plant binary vector for transiently expressing a single guide RNA (sgRNA) together with Cas9.

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pRGE31 Sequence and MappRGE31.dna
Map and Sequence File   
Sequence Author:  Yang Lab / Addgene #50929
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 SV40 NLS ATG BsaI (3819) BsaI (3839) polIII terminator attB1 pRGE31 9188 bp
BsaI  (3819)
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  (3839)
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.
Cas9
4998 .. 9098  =  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
4998 .. 9098  =  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
AmpR
1403 .. 2263  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1403 .. 1471  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1403 .. 2263  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1472 .. 2263  =  792 bp
   263 amino acids  =  29.0 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1403 .. 2263  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2434 .. 3022  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2434 .. 3022  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
561 .. 1016  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
561 .. 1016  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
OsU3 promoter
3443 .. 3823  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
OsU3 promoter
3443 .. 3823  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
CaMV 35S promoter
4481 .. 4826  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
4481 .. 4826  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
71 .. 323  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
71 .. 323  =  253 bp
nopaline synthase terminator and poly(A) signal
AmpR promoter
1298 .. 1402  =  105 bp
AmpR promoter
1298 .. 1402  =  105 bp
gRNA scaffold
3840 .. 3915  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
3840 .. 3915  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
3xFLAG
4881 .. 4946  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
3xFLAG
4881 .. 4946  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
nucleoplasmin NLS
9099 .. 9146  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
9099 .. 9146  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
attB1
4848 .. 4872  =  25 bp
recombination site for the Gateway® BP reaction
attB1
4848 .. 4872  =  25 bp
recombination site for the Gateway® BP reaction
attB2
9179 .. 15  =  25 bp
recombination site for the Gateway® BP reaction
attB2
9179 .. 15  =  25 bp
recombination site for the Gateway® BP reaction
SV40 NLS
4953 .. 4973  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
4953 .. 4973  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
3916 .. 3921  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3916 .. 3921  =  6 bp
RNA polymerase III transcription terminator
ATG
4878 .. 4880  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4878 .. 4880  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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