3xFLAG-dCas9 pMXs-puro

Retroviral vector with a puromycin resistance marker, encoding FLAG®-tagged dCas9 for engineered ChIP (enChIP) purification of specific genomic regions.

Sequence Author: Fujii Lab / Addgene #51240

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ATG 3xFLAG-dCas9/pMXs-puro 10,003 bp
ATG
1948 .. 1950  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1948 .. 1950  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
1951 .. 2016  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
1951 .. 2016  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
dCas9
2020 .. 6123  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   2020 .. 2046  =  27 bp
   9 amino acids  =  1.1 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2020 .. 6123  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   2047 .. 2049  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2020 .. 6123  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   2050 .. 4536  =  2487 bp
   829 amino acids  =  96.3 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2020 .. 6123  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   4537 .. 4539  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2020 .. 6123  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   4540 .. 6123  =  1584 bp
   528 amino acids  =  60.9 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2020 .. 6123  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
SV40 NLS
6136 .. 6156  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6136 .. 6156  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
AmpR
9030 .. 9890  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   9030 .. 9821  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9030 .. 9890  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   9822 .. 9890  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9030 .. 9890  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
6682 .. 7281  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
6682 .. 7281  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
LTR
22 .. 615  =  594 bp
long terminal repeat from Moloney murine leukemia virus
LTR
22 .. 615  =  594 bp
long terminal repeat from Moloney murine leukemia virus
ori
8278 .. 8866  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
8278 .. 8866  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
LTR
7508 .. 7977  =  470 bp
long terminal repeat from Moloney murine leukemia virus
LTR
7508 .. 7977  =  470 bp
long terminal repeat from Moloney murine leukemia virus
gag (truncated)
1094 .. 1510  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
1094 .. 1510  =  417 bp
truncated MMLV gag gene lacking the start codon
pol region
1520 .. 1894  =  375 bp
Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site
pol region
1520 .. 1894  =  375 bp
Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site
MMLV Ψ
678 .. 1035  =  358 bp
packaging signal of Moloney murine leukemia virus (MMLV)
MMLV Ψ
678 .. 1035  =  358 bp
packaging signal of Moloney murine leukemia virus (MMLV)
SV40 promoter
6343 .. 6672  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
6343 .. 6672  =  330 bp
SV40 enhancer and early promoter
AmpR promoter
9891 .. 9995  =  105 bp
AmpR promoter
9891 .. 9995  =  105 bp
SV40 ori
6523 .. 6658  =  136 bp
SV40 origin of replication
SV40 ori
6523 .. 6658  =  136 bp
SV40 origin of replication
ORF:  1948 .. 6159  =  4212 bp
ORF:  1403 amino acids  =  162.6 kDa
ORF:  6682 .. 7281  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  9160 .. 9426  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1577 .. 1912  =  336 bp
ORF:  111 amino acids  =  12.3 kDa
ORF:  6086 .. 6355  =  270 bp
ORF:  89 amino acids  =  10.0 kDa
ORF:  6291 .. 6620  =  330 bp
ORF:  109 amino acids  =  12.4 kDa
ORF:  1942 .. 2499  =  558 bp
ORF:  185 amino acids  =  20.7 kDa
ORF:  3424 .. 3942  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  4006 .. 4281  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  5533 .. 6066  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  6658 .. 7332  =  675 bp
ORF:  224 amino acids  =  22.9 kDa
ORF:  9787 .. 95  =  312 bp
ORF:  103 amino acids  =  12.0 kDa
ORF:  9030 .. 9890  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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