pAC2-dual-dCas9VP48-sgExpression

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BbsI (245) AAV2 ITR SV40 NLS BbsI (267) CMV enhancer chicken β-actin promoter ATG SV40 NLS pAC2-dual-dCas9VP48-sgExpression 8565 bp
BbsI  (245)
2 sites
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbsI  (267)
2 sites
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
ATG
1251 .. 1253  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1251 .. 1253  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
HA
1254 .. 1280  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
1254 .. 1280  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
SV40 NLS
1284 .. 1304  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1284 .. 1304  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
dCas9
1314 .. 5414  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 1:  
   1314 .. 1337  =  24 bp
   8 amino acids  =  923.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
1314 .. 5414  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 2:  
   1338 .. 1340  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
1314 .. 5414  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 3:  
   1341 .. 3827  =  2487 bp
   829 amino acids  =  96.3 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
1314 .. 5414  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 4:  
   3828 .. 3830  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
1314 .. 5414  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 5:  
   3831 .. 5414  =  1584 bp
   528 amino acids  =  60.9 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
1314 .. 5414  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
SV40 NLS
5418 .. 5438  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5418 .. 5438  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
VP48
5457 .. 5567  =  111 bp
37 amino acids  =  4.1 kDa
Product: 3 tandem repeats of the minimal activation domain of herpes simplex virus VP16 (Cheng et al., 2013)
VP48
5457 .. 5567  =  111 bp
37 amino acids  =  4.1 kDa
Product: 3 tandem repeats of the minimal activation domain of herpes simplex virus VP16 (Cheng et al., 2013)
AmpR
6885 .. 7745  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6885 .. 6953  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6885 .. 7745  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6954 .. 7745  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6885 .. 7745  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7916 .. 8504  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7916 .. 8504  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6043 .. 6498  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6043 .. 6498  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
440 .. 725  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
CMV enhancer
440 .. 725  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
chicken β-actin promoter
727 .. 1004  =  278 bp
chicken β-actin promoter
727 .. 1004  =  278 bp
U6 promoter
1 .. 249  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
U6 promoter
1 .. 249  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
hybrid intron
1005 .. 1233  =  229 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
hybrid intron
1005 .. 1233  =  229 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
bGH poly(A) signal
5612 .. 5819  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5612 .. 5819  =  208 bp
bovine growth hormone polyadenylation signal
AAV2 ITR
5828 .. 5968  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
5828 .. 5968  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
6780 .. 6884  =  105 bp
AmpR promoter
6780 .. 6884  =  105 bp
gRNA scaffold
268 .. 343  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
268 .. 343  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
polIII terminator
344 .. 349  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
344 .. 349  =  6 bp
RNA polymerase III transcription terminator
ORF:  5842 .. 6066  =  225 bp
ORF:  74 amino acids  =  8.3 kDa
ORF:  635 .. 982  =  348 bp
ORF:  115 amino acids  =  12.1 kDa
ORF:  825 .. 1136  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  1251 .. 5576  =  4326 bp
ORF:  1441 amino acids  =  166.4 kDa
ORF:  6885 .. 7745  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  5742 .. 6023  =  282 bp
ORF:  93 amino acids  =  10.4 kDa
ORF:  2660 .. 2953  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  3224 .. 3673  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  7349 .. 7615  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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