pX334

Zhang lab vector for expressing a Cas9 nickase mutant, a tracrRNA, and a guide sequence. Also known as pX334-U6-DR-BB-DR-Cbh-NLS-hSpCas9n(D10A)-NLS-H1-shorttracr-PGK-puro.

Sequence Author: Zhang Lab / Addgene #42333

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: CRISPR Plasmids | More Plasmid Sets
No matches
DR polIII terminator tracrRNA BbsI (283) BbsI (305) DR CMV enhancer chicken β-actin promoter ATG HA SV40 NLS pX334 10,152 bp
BbsI  (283)
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  (305)
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
1200 .. 1202  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1200 .. 1202  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
HA
1203 .. 1229  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
1203 .. 1229  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
SV40 NLS
1233 .. 1253  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1233 .. 1253  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9(D10A)
1263 .. 5363  =  4101 bp
1367 amino acids  =  158.3 kDa
3 segments
   Segment 1:  
   1263 .. 1286  =  24 bp
   8 amino acids  =  923.1 Da
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
Cas9(D10A)
1263 .. 5363  =  4101 bp
1367 amino acids  =  158.3 kDa
3 segments
   Segment 2:  
   1287 .. 1289  =  3 bp
   1 amino acid  =  89.1 Da
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
Cas9(D10A)
1263 .. 5363  =  4101 bp
1367 amino acids  =  158.3 kDa
3 segments
   Segment 3:  
   1290 .. 5363  =  4074 bp
   1358 amino acids  =  157.3 kDa
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
Cas9(D10A)
1263 .. 5363  =  4101 bp
1367 amino acids  =  158.3 kDa
3 segments
Product: nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain
SV40 NLS
5367 .. 5387  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5367 .. 5387  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
AmpR
8472 .. 9332  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8472 .. 8540  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8472 .. 9332  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8541 .. 9332  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8472 .. 9332  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
6758 .. 7357  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
6758 .. 7357  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
9503 .. 10,091  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9503 .. 10,091  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
6219 .. 6718  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
6219 .. 6718  =  500 bp
mouse phosphoglycerate kinase 1 promoter
f1 ori
7630 .. 8085  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
7630 .. 8085  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
390 .. 675  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
CMV enhancer
390 .. 675  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
chicken β-actin promoter
677 .. 954  =  278 bp
chicken β-actin promoter
677 .. 954  =  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
955 .. 1182  =  228 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
hybrid intron
955 .. 1182  =  228 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
H1 promoter
5651 .. 5865  =  215 bp
human H1 RNA promoter
H1 promoter
5651 .. 5865  =  215 bp
human H1 RNA promoter
bGH poly(A) signal
5430 .. 5637  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5430 .. 5637  =  208 bp
bovine growth hormone polyadenylation signal
AmpR promoter
8367 .. 8471  =  105 bp
AmpR promoter
8367 .. 8471  =  105 bp
tracrRNA
5876 .. 5954  =  79 bp
trans-activating CRISPR RNA for the Streptococcus pyogenes CRISPR/Cas9 system
tracrRNA
5876 .. 5954  =  79 bp
trans-activating CRISPR RNA for the Streptococcus pyogenes CRISPR/Cas9 system
DR
304 .. 339  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR system
DR
304 .. 339  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR system
DR
252 .. 287  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR system
DR
252 .. 287  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR system
polIII terminator
340 .. 344  =  5 bp
RNA polymerase III transcription terminator
polIII terminator
340 .. 344  =  5 bp
RNA polymerase III transcription terminator
polIII terminator
5955 .. 5959  =  5 bp
RNA polymerase III transcription terminator
polIII terminator
5955 .. 5959  =  5 bp
RNA polymerase III transcription terminator
ORF:  775 .. 1026  =  252 bp
ORF:  83 amino acids  =  8.7 kDa
ORF:  6499 .. 7446  =  948 bp
ORF:  315 amino acids  =  34.8 kDa
ORF:  6758 .. 7357  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  327 .. 569  =  243 bp
ORF:  80 amino acids  =  8.8 kDa
ORF:  585 .. 932  =  348 bp
ORF:  115 amino acids  =  12.1 kDa
ORF:  1200 .. 5399  =  4200 bp
ORF:  1399 amino acids  =  162.0 kDa
ORF:  8472 .. 9332  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1179 .. 3779  =  2601 bp
ORF:  866 amino acids  =  91.2 kDa
ORF:  7290 .. 7610  =  321 bp
ORF:  106 amino acids  =  12.3 kDa
ORF:  2609 .. 2902  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  3173 .. 3622  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  5420 .. 5665  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  6731 .. 7408  =  678 bp
ORF:  225 amino acids  =  23.1 kDa
ORF:  8936 .. 9202  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
Click here to try SnapGene

Download pX334.dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps

The maps, notes, and annotations in the zip file on this page are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as ’’www.snapgene.com/resources’’. Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Discover the most user-friendly molecular biology experience.