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Plasmid Files

pRGEB32

Plant vector for expressing a single guide RNA (sgRNA) with the rice U3 promoter and Cas9 with the rice ubiquitin promoter.

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pRGEB32 Sequence and MappRGEB32.dna
Map and Sequence File   
Sequence Author:  Yang Lab / Addgene #63159
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 SV40 NLS ATG polIII terminator BsaI (9652) BsaI (9632) RB T-DNA repeat pRGEB32 15,905 bp
BsaI  (9652)
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  (9632)
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
11,715 .. 15,815  =  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
11,715 .. 15,815  =  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
Ubi promoter
9809 .. 11,518  =  1710 bp
promoter for the rice (Oryza sativa) polyubiquitin
gene RUBQ2
Ubi promoter
9809 .. 11,518  =  1710 bp
promoter for the rice (Oryza sativa) polyubiquitin
gene RUBQ2
pVS1 RepA
2955 .. 4022  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
pVS1 RepA
2955 .. 4022  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
HygR
7162 .. 8187  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
7162 .. 8187  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
KanR
5627 .. 6421  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
5627 .. 6421  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
8255 .. 8932  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
8255 .. 8932  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
1892 .. 2521  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
pVS1 StaA
1892 .. 2521  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
ori
4952 .. 5540  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4952 .. 5540  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
OsU3 promoter
9256 .. 9636  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
OsU3 promoter
9256 .. 9636  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
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
pVS1 oriV
4088 .. 4282  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
4088 .. 4282  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
6948 .. 7122  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6948 .. 7122  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
4626 .. 4766  =  141 bp
basis of mobility region from pBR322
bom
4626 .. 4766  =  141 bp
basis of mobility region from pBR322
gRNA scaffold
9653 .. 9728  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
9653 .. 9728  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
3xFLAG
11,598 .. 11,663  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
3xFLAG
11,598 .. 11,663  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
nucleoplasmin NLS
15,816 .. 15,863  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
15,816 .. 15,863  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
RB T-DNA repeat
568 .. 592  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
568 .. 592  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6846 .. 6870  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6846 .. 6870  =  25 bp
left border repeat from nopaline C58 T-DNA
attB1
11,529 .. 11,553  =  25 bp
recombination site for the Gateway® BP reaction
attB1
11,529 .. 11,553  =  25 bp
recombination site for the Gateway® BP reaction
attB2
15,896 .. 15  =  25 bp
recombination site for the Gateway® BP reaction
attB2
15,896 .. 15  =  25 bp
recombination site for the Gateway® BP reaction
SV40 NLS
11,670 .. 11,690  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
11,670 .. 11,690  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
9729 .. 9734  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
9729 .. 9734  =  6 bp
RNA polymerase III transcription terminator
ATG
11,595 .. 11,597  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
11,595 .. 11,597  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
intron
10,557 .. 11,518  =  962 bp
intron upstream of the coding sequence for the rice
polyubiquitin gene RUBQ2
intron
10,557 .. 11,518  =  962 bp
intron upstream of the coding sequence for the rice
polyubiquitin gene RUBQ2
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