PX856

Zhang lab split-Cas9 plasmid for expressing FKBP fused to a C-terminal fragment of catalytically inactive dCas9 and the VP64 transcriptional activation domain.

Sequence Author: Zhang Lab / Addgene #62888

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BbsI (245) BbsI (267) CMV enhancer hybrid intron ATG PX856 7351 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
SV40 NLS
1257 .. 1277  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1257 .. 1277  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
FKBP
1302 .. 1622  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
synthetic gene with alternative codons
FKBP
1302 .. 1622  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
synthetic gene with alternative codons
dCas9(C)
1668 .. 4166  =  2499 bp
833 amino acids  =  96.4 kDa
3 segments
   Segment 1:  
   1668 .. 2648  =  981 bp
   327 amino acids  =  38.1 kDa
Product: C-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 carrying the N863A mutation (Zetsche et al., 2015)
dCas9(C)
1668 .. 4166  =  2499 bp
833 amino acids  =  96.4 kDa
3 segments
   Segment 2:  
   2649 .. 2651  =  3 bp
   1 amino acid  =  89.1 Da
Product: C-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 carrying the N863A mutation (Zetsche et al., 2015)
dCas9(C)
1668 .. 4166  =  2499 bp
833 amino acids  =  96.4 kDa
3 segments
   Segment 3:  
   2652 .. 4166  =  1515 bp
   505 amino acids  =  58.3 kDa
Product: C-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 carrying the N863A mutation (Zetsche et al., 2015)
dCas9(C)
1668 .. 4166  =  2499 bp
833 amino acids  =  96.4 kDa
3 segments
Product: C-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 carrying the N863A mutation (Zetsche et al., 2015)
SV40 NLS
4170 .. 4190  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4170 .. 4190  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
VP64
4209 .. 4358  =  150 bp
50 amino acids  =  5.5 kDa
Product: tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)
VP64
4209 .. 4358  =  150 bp
50 amino acids  =  5.5 kDa
Product: tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)
AmpR
5671 .. 6531  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5671 .. 5739  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5671 .. 6531  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5740 .. 6531  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5671 .. 6531  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
6702 .. 7290  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6702 .. 7290  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4829 .. 5284  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4829 .. 5284  =  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 .. 241  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
1 .. 241  =  241 bp
RNA polymerase III promoter for human U6 snRNA
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
4398 .. 4605  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
4398 .. 4605  =  208 bp
bovine growth hormone polyadenylation signal
AAV2 ITR
4614 .. 4754  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
4614 .. 4754  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
5566 .. 5670  =  105 bp
AmpR promoter
5566 .. 5670  =  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 transcription terminator
polIII terminator
344 .. 349  =  6 bp
RNA polymerase transcription terminator
ORF:  1189 .. 1467  =  279 bp
ORF:  92 amino acids  =  10.3 kDa
ORF:  5671 .. 6531  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  635 .. 982  =  348 bp
ORF:  115 amino acids  =  12.1 kDa
ORF:  4628 .. 4852  =  225 bp
ORF:  74 amino acids  =  8.3 kDa
ORF:  825 .. 1136  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  1251 .. 4367  =  3117 bp
ORF:  1038 amino acids  =  118.2 kDa
ORF:  1976 .. 2425  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  4528 .. 4809  =  282 bp
ORF:  93 amino acids  =  10.4 kDa
ORF:  1230 .. 2582  =  1353 bp
ORF:  450 amino acids  =  47.3 kDa
ORF:  6135 .. 6401  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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