pCLIP-Cas9-Nuclease-TRE3G-Puro

Lentiviral CRISPR vector encoding Cas9 and a puromycin resistance marker expressed from a Tet-responsive promoter.

Sequence Author: transOMIC

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FLAG CMV enhancer HIV-1 Ψ pCLIP-Cas9-Nuclease-TRE3G-Puro 10,794 bp
Cas9
2640 .. 6743  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
2640 .. 6743  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
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.
PuroR
6882 .. 7478  =  597 bp
198 amino acids  =  21.4 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
6882 .. 7478  =  597 bp
198 amino acids  =  21.4 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
AmpR
9799 .. 10,659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   9799 .. 10,590  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9799 .. 10,659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,591 .. 10,659  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9799 .. 10,659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
7503 .. 8091  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
7503 .. 8091  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
9043 .. 9628  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9043 .. 9628  =  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
8625 .. 8954  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
8625 .. 8954  =  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)
8296 .. 8529  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
8296 .. 8529  =  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
10,660 .. 10,764  =  105 bp
AmpR promoter
10,660 .. 10,764  =  105 bp
ORF:  2275 .. 2541  =  267 bp
ORF:  88 amino acids  =  10.5 kDa
ORF:  6859 .. 7173  =  315 bp
ORF:  104 amino acids  =  10.9 kDa
ORF:  7606 .. 8118  =  513 bp
ORF:  170 amino acids  =  18.0 kDa
ORF:  9929 .. 10,195  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1215 .. 2066  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  2640 .. 7478  =  4839 bp
ORF:  1612 amino acids  =  184.7 kDa
ORF:  7593 .. 8375  =  783 bp
ORF:  260 amino acids  =  28.8 kDa
ORF:  9799 .. 10,659  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2610 .. 5159  =  2550 bp
ORF:  849 amino acids  =  89.1 kDa
ORF:  10,556 .. 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
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