pK7WGF2_hCas9

Plant vector for expressing human codon-optimized Cas9 with an N-terminal enhanced GFP (EGFP) tag.

Sequence Author: Kamoun Lab / Addgene #46965

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attB1 SV40 NLS LB T-DNA repeat pK7WGF2::hCas9 14,400 bp
EGFP
13,633 .. 14,349  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   13,633 .. 13,635  =  3 bp
   1 amino acid  =  149.2 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
13,633 .. 14,349  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   13,636 .. 13,638  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
13,633 .. 14,349  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   13,639 .. 14,349  =  711 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
13,633 .. 14,349  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
Product: enhanced GFP
mammalian codon-optimized
Cas9
1 .. 4104  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
1 .. 4104  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SV40 NLS
4117 .. 4137  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4117 .. 4137  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
pVS1 RepA
9049 .. 10,116  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 RepA
9049 .. 10,116  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
NeoR/KanR
4765 .. 5559  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
4765 .. 5559  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
SmR
6496 .. 7287  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase (Murphy, 1985)
confers resistance to spectinomycin and streptomycin
SmR
6496 .. 7287  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase (Murphy, 1985)
confers resistance to spectinomycin and streptomycin
pVS1 StaA
10,550 .. 11,179  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 StaA
10,550 .. 11,179  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
ori
7531 .. 8119  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7531 .. 8119  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CaMV 35S promoter
13,196 .. 13,540  =  345 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
13,196 .. 13,540  =  345 bp
strong constitutive promoter from cauliflower mosaic virus
NOS terminator
5599 .. 5851  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
5599 .. 5851  =  253 bp
nopaline synthase terminator and poly(A) signal
pVS1 oriV
8789 .. 8983  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 oriV
8789 .. 8983  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
NOS promoter
4552 .. 4731  =  180 bp
nopaline synthase promoter
NOS promoter
4552 .. 4731  =  180 bp
nopaline synthase promoter
bom
8305 .. 8445  =  141 bp
basis of mobility region from pBR322
bom
8305 .. 8445  =  141 bp
basis of mobility region from pBR322
attB2
4156 .. 4180  =  25 bp
recombination site for the Gateway® BP reaction
attB2
4156 .. 4180  =  25 bp
recombination site for the Gateway® BP reaction
LB T-DNA repeat
5944 .. 5968  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
5944 .. 5968  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
12,479 .. 12,503  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
12,479 .. 12,503  =  25 bp
right border repeat from nopaline C58 T-DNA
attB1
14,356 .. 14,380  =  25 bp
recombination site for the Gateway® BP reaction
attB1
14,356 .. 14,380  =  25 bp
recombination site for the Gateway® BP reaction
ORF:  4765 .. 5559  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  6355 .. 7287  =  933 bp
ORF:  310 amino acids  =  34.5 kDa
ORF:  9682 .. 10,275  =  594 bp
ORF:  197 amino acids  =  22.2 kDa
ORF:  11,782 .. 12,132  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  12,703 .. 12,966  =  264 bp
ORF:  87 amino acids  =  10.4 kDa
ORF:  13,468 .. 4140  =  5073 bp
ORF:  1690 amino acids  =  194.9 kDa
ORF:  4937 .. 5323  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  10,019 .. 10,282  =  264 bp
ORF:  87 amino acids  =  9.1 kDa
ORF:  11,441 .. 11,713  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  11,127 .. 11,441  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  12,624 .. 13,442  =  819 bp
ORF:  272 amino acids  =  30.9 kDa
ORF:  13,507 .. 480  =  1374 bp
ORF:  457 amino acids  =  47.9 kDa
ORF:  1405 .. 1923  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  1987 .. 2520  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
ORF:  3514 .. 4047  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  5890 .. 6231  =  342 bp
ORF:  113 amino acids  =  12.1 kDa
ORF:  6535 .. 6918  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  9049 .. 10,155  =  1107 bp
ORF:  368 amino acids  =  41.2 kDa
ORF:  14,283 .. 143  =  261 bp
ORF:  86 amino acids  =  9.7 kDa
ORF:  6528 .. 6785  =  258 bp
ORF:  85 amino acids  =  9.1 kDa
ORF:  6795 .. 7073  =  279 bp
ORF:  92 amino acids  =  9.4 kDa
ORF:  11,478 .. 12,164  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  12,651 .. 12,950  =  300 bp
ORF:  99 amino acids  =  11.2 kDa
ORF:  6101 .. 6343  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  10,550 .. 11,179  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
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