pHAGE-TO-nmdCas9-3XmCherry

Lentiviral vector for labeling chromosomal loci by tetracycline-regulatable expression of catalytically dead Neisseria meningitidis dCas9 fused to three copies of mCherry.

Sequence Author: Pederson Lab / Addgene #64110

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CMV promoter tet operator tet operator ATG pHAGE-TO-nmdCas9-3XmCherry 12,104 bp
ATG
2980 .. 2982  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2980 .. 2982  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
2986 .. 3006  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2986 .. 3006  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
   Segment 1:  
   3019 .. 3060  =  42 bp
   14 amino acids  =  1.5 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
   Segment 2:  
   3061 .. 3063  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
   Segment 3:  
   3064 .. 4773  =  1710 bp
   570 amino acids  =  65.1 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
   Segment 4:  
   4774 .. 4779  =  6 bp
   2 amino acids  =  160.2 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
   Segment 5:  
   4780 .. 4845  =  66 bp
   22 amino acids  =  2.5 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
   Segment 6:  
   4846 .. 4848  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
   Segment 7:  
   4849 .. 6261  =  1413 bp
   471 amino acids  =  54.7 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
NmdCas9
3019 .. 6261  =  3243 bp
1081 amino acids  =  124.0 kDa
7 segments
Product: catalytically dead mutant of the Cas9 endonuclease from the Neisseria meningitidis Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D16A, D587A, H588A, and N611A
SV40 NLS
6280 .. 6300  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6280 .. 6300  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
mCherry
6313 .. 7020  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
6313 .. 7020  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
7039 .. 7746  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
7039 .. 7746  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
7765 .. 8472  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
7765 .. 8472  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
AmpR
9700 .. 10,560  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   9700 .. 9768  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9700 .. 10,560  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   9769 .. 10,560  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9700 .. 10,560  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
LTR
1 .. 634  =  634 bp
long terminal repeat (LTR) from HIV-1
LTR
1 .. 634  =  634 bp
long terminal repeat (LTR) from HIV-1
WPRE
8494 .. 9082  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
8494 .. 9082  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
10,731 .. 11,319  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
10,731 .. 11,319  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
2299 .. 2602  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2299 .. 2602  =  304 bp
human cytomegalovirus immediate early enhancer
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
LTR (ΔU3)
9157 .. 9390  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
LTR (ΔU3)
9157 .. 9390  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
CMV promoter
2603 .. 2806  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2603 .. 2806  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2058 .. 2175  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2058 .. 2175  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
9595 .. 9699  =  105 bp
AmpR promoter
9595 .. 9699  =  105 bp
tet operator
2808 .. 2826  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2808 .. 2826  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2829 .. 2847  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2829 .. 2847  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  2980 .. 8481  =  5502 bp
ORF:  1833 amino acids  =  208.5 kDa
ORF:  8584 .. 9096  =  513 bp
ORF:  170 amino acids  =  18.6 kDa
ORF:  9700 .. 10,560  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1181 .. 2107  =  927 bp
ORF:  308 amino acids  =  35.4 kDa
ORF:  2762 .. 3130  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  8597 .. 9130  =  534 bp
ORF:  177 amino acids  =  18.8 kDa
ORF:  1881 .. 2129  =  249 bp
ORF:  82 amino acids  =  9.0 kDa
ORF:  4071 .. 4307  =  237 bp
ORF:  78 amino acids  =  9.1 kDa
ORF:  6069 .. 6380  =  312 bp
ORF:  103 amino acids  =  12.2 kDa
ORF:  10,164 .. 10,430  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
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Download pHAGE-TO-nmdCas9-3XmCherry.dna file

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