HBT-pcoCas9

Plasmid for transient expression of plant codon-optimized Cas9 in plant cells. Use together with pUC119-gRNA.

Sequence Author: Sheen Lab / Addgene #52254

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ATG HBT-pcoCas9 7940 bp
ATG
3240 .. 3242  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3240 .. 3242  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
2xFLAG
3243 .. 3290  =  48 bp
16 amino acids  =  2.0 kDa
Product: two tandem FLAG® epitope tags
2xFLAG
3243 .. 3290  =  48 bp
16 amino acids  =  2.0 kDa
Product: two tandem FLAG® epitope tags
SV40 NLS
3297 .. 3317  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
3297 .. 3317  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
pcoCas9
3342 .. 7631  =  4290 bp
1367 amino acids  =  158.3 kDa
2 segments  =  4101 bp
Product: Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
plant codon-optimized Cas9 gene containing the potato IV2 intron
pcoCas9
3342 .. 7631  =  4290 bp
1367 amino acids  =  158.3 kDa
2 segments  =  4101 bp
   Segment 1:  
   3342 .. 4778  =  1437 bp
   479 amino acids  =  55.4 kDa
Product: Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
plant codon-optimized Cas9 gene containing the potato IV2 intron
pcoCas9
3342 .. 7631  =  4290 bp
1367 amino acids  =  158.3 kDa
2 segments  =  4101 bp
   Segment 2:  
   4968 .. 7631  =  2664 bp
   888 amino acids  =  102.9 kDa
Product: Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
plant codon-optimized Cas9 gene containing the potato IV2 intron
pcoCas9
3342 .. 7631  =  4290 bp
1367 amino acids  =  158.3 kDa
2 segments  =  4101 bp
Product: Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
plant codon-optimized Cas9 gene containing the potato IV2 intron
nucleoplasmin NLS
7632 .. 7679  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
7632 .. 7679  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
AmpR
1200 .. 2060  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1200 .. 1991  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1200 .. 2060  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1992 .. 2060  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1200 .. 2060  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
441 .. 1029  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
441 .. 1029  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
35SPPDK promoter
2722 .. 3231  =  510 bp
3 segments
   Segment 1:  CaMV 35S enhancer  
   2722 .. 3020  =  299 bp
hybrid promoter consisting of the cauliflower mosaic virus 35S enhancer fused to the maize C4PPDK basal promoter (Yoo et al., 2007)
35SPPDK promoter
2722 .. 3231  =  510 bp
3 segments
   Segment 2:  
   3021 .. 3043  =  23 bp
hybrid promoter consisting of the cauliflower mosaic virus 35S enhancer fused to the maize C4PPDK basal promoter (Yoo et al., 2007)
35SPPDK promoter
2722 .. 3231  =  510 bp
3 segments
   Segment 3:  C4PPDK basal promoter  
   3044 .. 3231  =  188 bp
hybrid promoter consisting of the cauliflower mosaic virus 35S enhancer fused to the maize C4PPDK basal promoter (Yoo et al., 2007)
35SPPDK promoter
2722 .. 3231  =  510 bp
3 segments
hybrid promoter consisting of the cauliflower mosaic virus 35S enhancer fused to the maize C4PPDK basal promoter (Yoo et al., 2007)
NOS terminator
7688 .. 7940  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
7688 .. 7940  =  253 bp
nopaline synthase terminator and poly(A) signal
AmpR promoter
2061 .. 2165  =  105 bp
AmpR promoter
2061 .. 2165  =  105 bp
IV2 intron
4779 .. 4967  =  189 bp
modified second intron of the potato ST-LS1 gene (Vancanneyt et al., 1990)
IV2 intron
4779 .. 4967  =  189 bp
modified second intron of the potato ST-LS1 gene (Vancanneyt et al., 1990)
ORF:  1330 .. 1596  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2446 .. 2724  =  279 bp
ORF:  92 amino acids  =  10.9 kDa
ORF:  3253 .. 3531  =  279 bp
ORF:  92 amino acids  =  11.1 kDa
ORF:  3877 .. 4302  =  426 bp
ORF:  141 amino acids  =  16.4 kDa
ORF:  5584 .. 5871  =  288 bp
ORF:  95 amino acids  =  11.5 kDa
ORF:  7327 .. 7782  =  456 bp
ORF:  151 amino acids  =  17.4 kDa
ORF:  2994 .. 4802  =  1809 bp
ORF:  602 amino acids  =  69.2 kDa
ORF:  5010 .. 7682  =  2673 bp
ORF:  890 amino acids  =  103.1 kDa
ORF:  1200 .. 2060  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3108 .. 4823  =  1716 bp
ORF:  571 amino acids  =  64.1 kDa
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Download HBT-pcoCas9.dna file

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