pSLQ1658-dCas9-EGFP

Retroviral vector encoding catalytically inactive dCas9 fused to enhanced GFP (EGFP), for visualizing genomic loci using a small guide RNA (sgRNA).

Sequence Author: Huang

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ATG pSLQ1658-dCas9-EGFP 11,177 bp
ATG
1423 .. 1425  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1423 .. 1425  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
1432 .. 1452  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1432 .. 1452  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
dCas9
1453 .. 5553  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 1:  
   1453 .. 1476  =  24 bp
   8 amino acids  =  923.1 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
1453 .. 5553  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 2:  
   1477 .. 1479  =  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
1453 .. 5553  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 3:  
   1480 .. 3966  =  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
1453 .. 5553  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 4:  
   3967 .. 3969  =  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
1453 .. 5553  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 5:  
   3970 .. 5553  =  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
1453 .. 5553  =  4101 bp
1367 amino acids  =  158.2 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
5563 .. 5583  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5563 .. 5583  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
EGFP
5593 .. 6309  =  717 bp
238 amino acids  =  26.8 kDa
2 segments
   Segment 1:  1a  
   5593 .. 5595  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
5593 .. 6309  =  717 bp
238 amino acids  =  26.8 kDa
2 segments
   Segment 2:  
   5596 .. 6309  =  714 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
5593 .. 6309  =  717 bp
238 amino acids  =  26.8 kDa
2 segments
Product: enhanced GFP
mammalian codon-optimized
AmpR
9383 .. 10,243  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   9383 .. 10,174  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9383 .. 10,243  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,175 .. 10,243  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9383 .. 10,243  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
6840 .. 7439  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
6840 .. 7439  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
8624 .. 9212  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
8624 .. 9212  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
5' LTR
11,176 .. 515  =  517 bp
5' long terminal repeat from murine embryonic stem cell virus
5' LTR
11,176 .. 515  =  517 bp
5' long terminal repeat from murine embryonic stem cell virus
3' LTR
7539 .. 8053  =  515 bp
3' long terminal repeat from murine embryonic stem cell virus
3' LTR
7539 .. 8053  =  515 bp
3' long terminal repeat from murine embryonic stem cell virus
PGK promoter
6320 .. 6819  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
6320 .. 6819  =  500 bp
mouse phosphoglycerate kinase 1 promoter
gag (truncated)
987 .. 1403  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
987 .. 1403  =  417 bp
truncated MMLV gag gene lacking the start codon
MESV Ψ
579 .. 920  =  342 bp
packaging signal of murine embryonic stem cell virus
MESV Ψ
579 .. 920  =  342 bp
packaging signal of murine embryonic stem cell virus
AmpR promoter
10,244 .. 10,348  =  105 bp
AmpR promoter
10,244 .. 10,348  =  105 bp
ORF:  1423 .. 6309  =  4887 bp
ORF:  1628 amino acids  =  187.5 kDa
ORF:  5300 .. 5584  =  285 bp
ORF:  94 amino acids  =  10.0 kDa
ORF:  6600 .. 7439  =  840 bp
ORF:  279 amino acids  =  30.1 kDa
ORF:  9513 .. 9779  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  10,629 .. 10,925  =  297 bp
ORF:  98 amino acids  =  11.7 kDa
ORF:  3363 .. 3731  =  369 bp
ORF:  122 amino acids  =  14.1 kDa
ORF:  6765 .. 7490  =  726 bp
ORF:  241 amino acids  =  24.8 kDa
ORF:  362 .. 637  =  276 bp
ORF:  91 amino acids  =  9.9 kDa
ORF:  674 .. 934  =  261 bp
ORF:  86 amino acids  =  9.5 kDa
ORF:  9383 .. 10,243  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3070 .. 3543  =  474 bp
ORF:  157 amino acids  =  17.4 kDa
ORF:  4855 .. 5241  =  387 bp
ORF:  128 amino acids  =  14.5 kDa
ORF:  10,693 .. 10,923  =  231 bp
ORF:  76 amino acids  =  7.5 kDa
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