pCLIP-Cas9(D10a)-Nickase-hCMV-tRFP

Lentiviral CRISPR vector encoding a Cas9 nickase mutant and the red fluorescent protein TurboRFP expressed from the human CMV promoter.

Sequence Author: transOMIC

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FLAG CMV enhancer HIV-1 Ψ pCLIP-Cas9-Nickase-hCMV-tRFP 11,158 bp
Cas9(D10A)
2905 .. 7008  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 1:  
   2905 .. 2931  =  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)
2905 .. 7008  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 2:  
   2932 .. 2934  =  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)
2905 .. 7008  =  4104 bp
1368 amino acids  =  158.4 kDa
3 segments
   Segment 3:  
   2935 .. 7008  =  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)
2905 .. 7008  =  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
7009 .. 7056  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
7009 .. 7056  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
FLAG
7057 .. 7080  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
7057 .. 7080  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
P2A
7090 .. 7146  =  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
7090 .. 7146  =  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.
TurboRFP
7147 .. 7842  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea quadricolor
mammalian codon-optimized
TurboRFP
7147 .. 7842  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea quadricolor
mammalian codon-optimized
AmpR
10,163 .. 11,023  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   10,163 .. 10,954  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,163 .. 11,023  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,955 .. 11,023  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,163 .. 11,023  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
7867 .. 8455  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
7867 .. 8455  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
9407 .. 9992  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9407 .. 9992  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
8989 .. 9318  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
8989 .. 9318  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
2238 .. 2541  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2238 .. 2541  =  304 bp
human cytomegalovirus immediate early enhancer
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
1333 .. 1566  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1333 .. 1566  =  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)
8660 .. 8893  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
8660 .. 8893  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
CMV promoter
2542 .. 2745  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2542 .. 2745  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
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
2057 .. 2174  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2057 .. 2174  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
11,024 .. 11,128  =  105 bp
AmpR promoter
11,024 .. 11,128  =  105 bp
ORF:  2905 .. 7842  =  4938 bp
ORF:  1645 amino acids  =  189.4 kDa
ORF:  7957 .. 8739  =  783 bp
ORF:  260 amino acids  =  28.8 kDa
ORF:  1211 .. 2062  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  7970 .. 8482  =  513 bp
ORF:  170 amino acids  =  18.0 kDa
ORF:  10,293 .. 10,559  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  10,920 .. 31  =  270 bp
ORF:  89 amino acids  =  10.4 kDa
ORF:  10,163 .. 11,023  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1126 .. 1506  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  2833 .. 5424  =  2592 bp
ORF:  863 amino acids  =  90.6 kDa
ORF:  4254 .. 4547  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  4818 .. 5267  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  7248 .. 7493  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
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