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Plasmid Files

pX260

Zhang lab vector for expressing Cas9, a tracrRNA, and a guide sequence. Also known as pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro.

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pX260 Sequence and MappX260.dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #42229
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 hybrid intron ATG 3xFLAG SV40 NLS tracrRNA polIII terminator BbsI (522) BbsI (544) polIII terminator pX260 10,203 bp
BbsI  (522)
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  (544)
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.
Cas9
5636 .. 9736  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
5636 .. 9736  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
AmpR
1652 .. 2512  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1652 .. 2443  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1652 .. 2512  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2444 .. 2512  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1652 .. 2512  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
3627 .. 4226  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3627 .. 4226  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
893 .. 1481  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
893 .. 1481  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
PGK promoter
4266 .. 4765  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
4266 .. 4765  =  500 bp
mouse phosphoglycerate kinase 1 promoter
f1 ori
2899 .. 3354  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2899 .. 3354  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
157 .. 442  =  286 bp
human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard
CMV enhancer
CMV enhancer
157 .. 442  =  286 bp
human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard
CMV enhancer
chicken β-actin promoter
10,102 .. 155  =  257 bp
contains a 21-bp deletion relative to the standard
chicken β-actin promoter
chicken β-actin promoter
10,102 .. 155  =  257 bp
contains a 21-bp deletion relative to the standard
chicken β-actin promoter
U6 promoter
591 .. 831  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
591 .. 831  =  241 bp
RNA polymerase III promoter for human U6 snRNA
hybrid intron
9874 .. 10,101  =  228 bp
hybrid between chicken β-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)
hybrid intron
9874 .. 10,101  =  228 bp
hybrid between chicken β-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)
H1 promoter
5119 .. 5333  =  215 bp
human H1 RNA promoter
H1 promoter
5119 .. 5333  =  215 bp
human H1 RNA promoter
bGH poly(A) signal
5347 .. 5554  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5347 .. 5554  =  208 bp
bovine growth hormone polyadenylation signal
AmpR promoter
2513 .. 2617  =  105 bp
AmpR promoter
2513 .. 2617  =  105 bp
tracrRNA
5030 .. 5108  =  79 bp
trans-activating CRISPR RNA for the Streptococcus
pyogenes
CRISPR/Cas9 system
tracrRNA
5030 .. 5108  =  79 bp
trans-activating CRISPR RNA for the Streptococcus
pyogenes
CRISPR/Cas9 system
3xFLAG
9788 .. 9853  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
3xFLAG
9788 .. 9853  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
nucleoplasmin NLS
5588 .. 5635  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
5588 .. 5635  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
DR
493 .. 528  =  36 bp
direct repeat for the Streptococcus pyogenes
CRISPR system
DR
493 .. 528  =  36 bp
direct repeat for the Streptococcus pyogenes
CRISPR system
DR
545 .. 580  =  36 bp
direct repeat for the Streptococcus pyogenes
CRISPR system
DR
545 .. 580  =  36 bp
direct repeat for the Streptococcus pyogenes
CRISPR system
SV40 NLS
9761 .. 9781  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
9761 .. 9781  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
488 .. 492  =  5 bp
RNA polymerase III transcription terminator
polIII terminator
488 .. 492  =  5 bp
RNA polymerase III transcription terminator
polIII terminator
5025 .. 5029  =  5 bp
RNA polymerase III transcription terminator
polIII terminator
5025 .. 5029  =  5 bp
RNA polymerase III transcription terminator
ATG
9854 .. 9856  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
9854 .. 9856  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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