pHAGE TRE dCas9-VP64

Lentiviral CRISPR activation vector for doxycycline-inducible expression of catalytically inactive dCas9 fused to the VP64 transcriptional activation domain.

Sequence Author: Maehr

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attB1 minimal CMV promoter attB2 pHAGE TRE dCas9-VP64 14,595 bp
dCas9
10,141 .. 14,244  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   10,141 .. 10,167  =  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
10,141 .. 14,244  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   10,168 .. 10,170  =  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
10,141 .. 14,244  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   10,171 .. 12,657  =  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
10,141 .. 14,244  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   12,658 .. 12,660  =  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
10,141 .. 14,244  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   12,661 .. 14,244  =  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
10,141 .. 14,244  =  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
14,257 .. 14,277  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
14,257 .. 14,277  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
3xHA
14,278 .. 14,367  =  90 bp
30 amino acids  =  3.5 kDa
Product: three tandem HA epitope tags
3xHA
14,278 .. 14,367  =  90 bp
30 amino acids  =  3.5 kDa
Product: three tandem HA epitope tags
VP64
14,389 .. 14,538  =  150 bp
50 amino acids  =  5.5 kDa
Product: tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)
VP64
14,389 .. 14,538  =  150 bp
50 amino acids  =  5.5 kDa
Product: tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)
UbC promoter
179 .. 1390  =  1212 bp
human ubiquitin C promoter
UbC promoter
179 .. 1390  =  1212 bp
human ubiquitin C promoter
AmpR
5002 .. 5862  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   5002 .. 5070  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5002 .. 5862  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   5071 .. 5862  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5002 .. 5862  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2823 .. 3617  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2823 .. 3617  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 1:  rTetR  
   1449 .. 2066  =  618 bp
   206 amino acids  =  23.2 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 2:  
   2067 .. 2072  =  6 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 3:  VP16 minimal transcriptional activation domain  
   2073 .. 2108  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 4:  
   2109 .. 2111  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 5:  VP16 minimal transcriptional activation domain  
   2112 .. 2147  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 6:  
   2148 .. 2150  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 7:  VP16 minimal transcriptional activation domain  
   2151 .. 2186  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 8:  
   2187 .. 2195  =  9 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
1449 .. 2195  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
LTR
7407 .. 8040  =  634 bp
long terminal repeat (LTR) from HIV-1
LTR
7407 .. 8040  =  634 bp
long terminal repeat (LTR) from HIV-1
WPRE
3796 .. 4384  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
3796 .. 4384  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6033 .. 6621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6033 .. 6621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
2233 .. 2807  =  575 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
2233 .. 2807  =  575 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 1:  tetO  
   9636 .. 9654  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 2:  
   9655 .. 9677  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 3:  tetO  
   9678 .. 9696  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 4:  
   9697 .. 9719  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 5:  tetO  
   9720 .. 9738  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 6:  
   9739 .. 9761  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 7:  tetO  
   9762 .. 9780  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 8:  
   9781 .. 9803  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 9:  tetO  
   9804 .. 9822  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 10:  
   9823 .. 9845  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 11:  tetO  
   9846 .. 9864  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 12:  
   9865 .. 9887  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
   Segment 13:  tetO  
   9888 .. 9906  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
9636 .. 9906  =  271 bp
13 segments
contains seven copies of the tetracycline operator tetO
LTR (ΔU3)
4459 .. 4692  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
LTR (ΔU3)
4459 .. 4692  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
RRE
8709 .. 8942  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
8709 .. 8942  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
HIV-1 Ψ
8087 .. 8212  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
8087 .. 8212  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
9464 .. 9581  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
9464 .. 9581  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
4897 .. 5001  =  105 bp
AmpR promoter
4897 .. 5001  =  105 bp
minimal CMV promoter
9939 .. 9977  =  39 bp
human cytomegalovirus (CMV) immediate early promoter
minimal CMV promoter
9939 .. 9977  =  39 bp
human cytomegalovirus (CMV) immediate early promoter
attB1
10,107 .. 10,131  =  25 bp
recombination site for the Gateway® BP reaction
attB1
10,107 .. 10,131  =  25 bp
recombination site for the Gateway® BP reaction
attB2
14,585 .. 14  =  25 bp
recombination site for the Gateway® BP reaction
attB2
14,585 .. 14  =  25 bp
recombination site for the Gateway® BP reaction
ORF:  1606 .. 1908  =  303 bp
ORF:  100 amino acids  =  11.1 kDa
ORF:  2995 .. 3381  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3886 .. 4398  =  513 bp
ORF:  170 amino acids  =  18.6 kDa
ORF:  5002 .. 5862  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  8587 .. 9513  =  927 bp
ORF:  308 amino acids  =  35.4 kDa
ORF:  10,141 .. 14,547  =  4407 bp
ORF:  1468 amino acids  =  169.2 kDa
ORF:  3899 .. 4432  =  534 bp
ORF:  177 amino acids  =  18.8 kDa
ORF:  14,207 .. 14,539  =  333 bp
ORF:  110 amino acids  =  12.5 kDa
ORF:  1449 .. 2195  =  747 bp
ORF:  248 amino acids  =  27.8 kDa
ORF:  2823 .. 3617  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  9444 .. 10,400  =  957 bp
ORF:  318 amino acids  =  37.3 kDa
ORF:  511 .. 804  =  294 bp
ORF:  97 amino acids  =  11.3 kDa
ORF:  2302 .. 2718  =  417 bp
ORF:  138 amino acids  =  15.2 kDa
ORF:  9619 .. 10,551  =  933 bp
ORF:  310 amino acids  =  33.5 kDa
ORF:  11,545 .. 12,063  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  12,127 .. 12,402  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  13,030 .. 14,187  =  1158 bp
ORF:  385 amino acids  =  43.7 kDa
ORF:  3132 .. 3386  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  5466 .. 5732  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  8421 .. 8882  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  1901 .. 2182  =  282 bp
ORF:  93 amino acids  =  9.5 kDa
ORF:  9287 .. 9535  =  249 bp
ORF:  82 amino acids  =  9.0 kDa
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Download pHAGE TRE dCas9-VP64.dna file

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