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Plasmid Files

pRGEB31

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

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pRGEB31 Sequence and MappRGEB31.dna
Map and Sequence File   
Sequence Author:  Yang Lab / Addgene #51295
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 BsaI (382) BsaI (402) polIII terminator attB1 ATG 3xFLAG SV40 NLS RB T-DNA repeat pRGEB31 15,037 bp
BsaI  (382)
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  (402)
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
1597 .. 5697  =  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
1597 .. 5697  =  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
pVS1 RepA
8742 .. 9809  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
pVS1 RepA
8742 .. 9809  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
HygR
12,949 .. 13,974  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
12,949 .. 13,974  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
KanR
11,414 .. 12,208  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
11,414 .. 12,208  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
14,042 .. 14,719  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
14,042 .. 14,719  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
7679 .. 8308  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
pVS1 StaA
7679 .. 8308  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas
plasmid pVS1 (Heeb et al., 2000)
ori
10,739 .. 11,327  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
10,739 .. 11,327  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
OsU3 promoter
6 .. 386  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
OsU3 promoter
6 .. 386  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
CaMV 35S promoter
1044 .. 1389  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
1044 .. 1389  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
5858 .. 6110  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
5858 .. 6110  =  253 bp
nopaline synthase terminator and poly(A) signal
pVS1 oriV
9875 .. 10,069  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
9875 .. 10,069  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
12,735 .. 12,909  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
12,735 .. 12,909  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
10,413 .. 10,553  =  141 bp
basis of mobility region from pBR322
bom
10,413 .. 10,553  =  141 bp
basis of mobility region from pBR322
gRNA scaffold
403 .. 478  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
403 .. 478  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
3xFLAG
1480 .. 1545  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
3xFLAG
1480 .. 1545  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
nucleoplasmin NLS
5698 .. 5745  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
5698 .. 5745  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
attB1
1411 .. 1435  =  25 bp
recombination site for the Gateway® BP reaction
attB1
1411 .. 1435  =  25 bp
recombination site for the Gateway® BP reaction
attB2
5778 .. 5802  =  25 bp
recombination site for the Gateway® BP reaction
attB2
5778 .. 5802  =  25 bp
recombination site for the Gateway® BP reaction
RB T-DNA repeat
6355 .. 6379  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
6355 .. 6379  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
12,633 .. 12,657  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
12,633 .. 12,657  =  25 bp
left border repeat from nopaline C58 T-DNA
SV40 NLS
1552 .. 1572  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
1552 .. 1572  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
479 .. 484  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
479 .. 484  =  6 bp
RNA polymerase III transcription terminator
ATG
1477 .. 1479  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1477 .. 1479  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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