pCLIP-Cas9(D10a)-Nickase-TRE3G-ZsGreen

Lentiviral CRISPR vector encoding a Cas9 nickase mutant and the green fluorescent protein ZsGreen expressed from a Tet-responsive promoter.

Sequence Author: transOMIC

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FLAG CMV enhancer HIV-1 Ψ pCLIP-Cas9-Nickase-TRE3G-ZsGreen 10,893 bp
Cas9(D10A)
2640 .. 6743  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 1:  
   2640 .. 2666  =  27 bp
   9 amino acids  =  1.1 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)
2640 .. 6743  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 2:  
   2667 .. 2669  =  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)
2640 .. 6743  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 3:  
   2670 .. 6743  =  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)
2640 .. 6743  =  4104 bp
1368 amino acids  =  158.4 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
nucleoplasmin NLS
6744 .. 6791  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
6744 .. 6791  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
FLAG
6792 .. 6815  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
6792 .. 6815  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
P2A
6825 .. 6881  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
P2A
6825 .. 6881  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
ZsGreen1
6882 .. 7577  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
6882 .. 7577  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
AmpR
9898 .. 10,758  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   9898 .. 10,689  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9898 .. 10,758  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,690 .. 10,758  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9898 .. 10,758  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
7602 .. 8190  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
7602 .. 8190  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
9142 .. 9727  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9142 .. 9727  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
TRE3G promoter
2244 .. 2622  =  379 bp
3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On® 3G
TRE3G promoter
2244 .. 2622  =  379 bp
3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On® 3G
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
8724 .. 9053  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
8724 .. 9053  =  330 bp
SV40 enhancer and early promoter
5' LTR (truncated)
411 .. 668  =  258 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
411 .. 668  =  258 bp
truncated 5' long terminal repeat (LTR) from HIV-1
RRE
1337 .. 1570  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1337 .. 1570  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
3' LTR (ΔU3)
8395 .. 8628  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
8395 .. 8628  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
715 .. 840  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
715 .. 840  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2062 .. 2177  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2062 .. 2177  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
10,759 .. 10,863  =  105 bp
AmpR promoter
10,759 .. 10,863  =  105 bp
ORF:  2275 .. 2541  =  267 bp
ORF:  88 amino acids  =  10.5 kDa
ORF:  7705 .. 8217  =  513 bp
ORF:  170 amino acids  =  18.0 kDa
ORF:  10,028 .. 10,294  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1215 .. 2066  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  2640 .. 7577  =  4938 bp
ORF:  1645 amino acids  =  189.4 kDa
ORF:  7692 .. 8474  =  783 bp
ORF:  260 amino acids  =  28.8 kDa
ORF:  9898 .. 10,758  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2610 .. 5159  =  2550 bp
ORF:  849 amino acids  =  89.1 kDa
ORF:  10,655 .. 31  =  270 bp
ORF:  89 amino acids  =  10.4 kDa
ORF:  1049 .. 1510  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  3989 .. 4282  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  4553 .. 5002  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  7259 .. 7552  =  294 bp
ORF:  97 amino acids  =  10.9 kDa
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Download pCLIP-Cas9(D10a)-Nickase-TRE3G-ZsGreen.dna file

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