PX854

Zhang lab split-Cas9 plasmid for expressing FKBP fused to a C-terminal fragment (amino acids 574-1368) of S. pyogenes Cas9.

Sequence Author: Zhang Lab / Addgene #62886

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U6 promoter BbsI (245) BbsI (267) polIII terminator ATG PX854 7087 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
FKBP
1302 .. 1622  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
Cas9(C)
1668 .. 4052  =  2385 bp
795 amino acids  =  92.0 kDa
Product: C-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)
Cas9(C)
1668 .. 4052  =  2385 bp
795 amino acids  =  92.0 kDa
Product: C-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)
nucleoplasmin NLS
4053 .. 4100  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
4053 .. 4100  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
AmpR
5407 .. 6267  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5407 .. 5475  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5407 .. 6267  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5476 .. 6267  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5407 .. 6267  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
6438 .. 7026  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6438 .. 7026  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4565 .. 5020  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4565 .. 5020  =  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
4134 .. 4341  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
4134 .. 4341  =  208 bp
bovine growth hormone polyadenylation signal
AAV2 ITR
4350 .. 4490  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
4350 .. 4490  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
5302 .. 5406  =  105 bp
AmpR promoter
5302 .. 5406  =  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:  1189 .. 1467  =  279 bp
ORF:  92 amino acids  =  10.3 kDa
ORF:  5407 .. 6267  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  635 .. 982  =  348 bp
ORF:  115 amino acids  =  12.1 kDa
ORF:  4364 .. 4588  =  225 bp
ORF:  74 amino acids  =  8.3 kDa
ORF:  825 .. 1136  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  1251 .. 4103  =  2853 bp
ORF:  950 amino acids  =  108.3 kDa
ORF:  1862 .. 2311  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  4264 .. 4545  =  282 bp
ORF:  93 amino acids  =  10.4 kDa
ORF:  1230 .. 2468  =  1239 bp
ORF:  412 amino acids  =  43.1 kDa
ORF:  5871 .. 6137  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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