SP-dCas9-VPR

Plasmid for mammalian cell expression of catalytically inactive Cas9m4 fused to the tripartite VP64-p65-Rta (VPR) transcriptional activator.

Sequence Author: Church Lab / Addgene #63798

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HomePlasmidsCRISPR PlasmidsSP-dCas9-VPR
attB2 SV40 NLS SP-dCas9-VPR 11,319 bp
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 1:  
   748 .. 774  =  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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 2:  
   775 .. 777  =  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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 3:  
   778 .. 3261  =  2484 bp
   828 amino acids  =  96.1 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 4:  
   3262 .. 3267  =  6 bp
   2 amino acids  =  160.2 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 5:  
   3268 .. 3333  =  66 bp
   22 amino acids  =  2.5 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 6:  
   3334 .. 3336  =  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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 7:  
   3337 .. 4851  =  1515 bp
   505 amino acids  =  58.3 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
SV40 NLS
4864 .. 4884  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4864 .. 4884  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
VP64
4975 .. 5124  =  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
4975 .. 5124  =  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)
SV40 NLS
5149 .. 5169  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5149 .. 5169  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
RelA (p65) AD
5599 .. 5955  =  357 bp
119 amino acids  =  12.4 kDa
Product: transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)
RelA (p65) AD
5599 .. 5955  =  357 bp
119 amino acids  =  12.4 kDa
Product: transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)
Rta AD
5974 .. 6543  =  570 bp
190 amino acids  =  20.6 kDa
Product: transcriptional activation domain from the human herpesvirus 4 (Epstein-Barr virus) replication and transcription activator Rta/BRLF1 (Hardwick et al., 1992; Chavez et al., 2015)
Rta AD
5974 .. 6543  =  570 bp
190 amino acids  =  20.6 kDa
Product: transcriptional activation domain from the human herpesvirus 4 (Epstein-Barr virus) replication and transcription activator Rta/BRLF1 (Hardwick et al., 1992; Chavez et al., 2015)
AmpR
10,138 .. 10,998  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   10,138 .. 10,929  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,138 .. 10,998  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,930 .. 10,998  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,138 .. 10,998  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
7837 .. 8631  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
7837 .. 8631  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
9379 .. 9967  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9379 .. 9967  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6999 .. 7427  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6999 .. 7427  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
50 .. 429  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
50 .. 429  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
7441 .. 7770  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
7441 .. 7770  =  330 bp
SV40 enhancer and early promoter
CMV promoter
430 .. 633  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
430 .. 633  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
8807 .. 8928  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8807 .. 8928  =  122 bp
SV40 polyadenylation signal
AmpR promoter
10,999 .. 11,103  =  105 bp
AmpR promoter
10,999 .. 11,103  =  105 bp
HSV TK poly(A) signal
6749 .. 6797  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
6749 .. 6797  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
attB1
4921 .. 4945  =  25 bp
recombination site for the Gateway® BP reaction
attB1
4921 .. 4945  =  25 bp
recombination site for the Gateway® BP reaction
attB2
6548 .. 6572  =  25 bp
recombination site for the Gateway® BP reaction
attB2
6548 .. 6572  =  25 bp
recombination site for the Gateway® BP reaction
SV40 ori
7621 .. 7756  =  136 bp
SV40 origin of replication
SV40 ori
7621 .. 7756  =  136 bp
SV40 origin of replication
ORF:  589 .. 6546  =  5958 bp
ORF:  1985 amino acids  =  225.0 kDa
ORF:  7837 .. 8631  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5891 .. 6175  =  285 bp
ORF:  94 amino acids  =  10.0 kDa
ORF:  8009 .. 8395  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  10,268 .. 10,534  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  724 .. 1227  =  504 bp
ORF:  167 amino acids  =  18.5 kDa
ORF:  2152 .. 2670  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  2734 .. 3009  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  4261 .. 4794  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  8146 .. 8400  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  10,138 .. 10,998  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  648 .. 890  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
ORF:  5529 .. 5762  =  234 bp
ORF:  77 amino acids  =  7.4 kDa
ORF:  6234 .. 6581  =  348 bp
ORF:  115 amino acids  =  12.7 kDa
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Individual Sequences & Maps

3xFLAG-dCas9 pCMV-7.1
pCas9_GFP
pCRISPRa-Enhancer
pME-Cas9-T2A-GFP
3xFLAG-dCas9 pMXs-neo
pCas9D10A_GFP
pCS2+hSpCas9
pMJ806
3xFLAG-dCas9 pMXs-puro
pCas-Guide
pCS2-Cas9
pMJ841
Cas9m4
pcDNA-dCas9
pCS2-nCas9n
pMJ920
Cas9m4-VP64
pcDNA-dCas9-p300 Core
pCW-Cas9
pMLM3705
CRISPRi library backbone
pcDNA-dCas9-p300 Core (D1399Y)
pdCas9
pMSCV-LTR-dCas9-p65AD-BFP
dCAS9-VP64_GFP
pcDNA-dCas9-VP64
pdCas9-bacteria
pMSCV-LTR-dCas9-VP64-BFP
eSpCas9(1.1)
pcDNA3.1-CibN-dCas9-CibN
pdCas9-humanized
pMZ374
GeneArt CRISPR Nuclease Vector - CD4 (linearized)
pCFD1-dU6-1gRNA
pdCas9 BFP-humanized
pNW3
GeneArt CRISPR Nuclease Vector - OFP (linearized)
pCFD2-dU6-2gRNA
pDCC6
pRGE31
gRNA_AAVS1-T1
pCFD3-dU6-3gRNA
pDD162 (Peft-3_Cas9 + Empty sgRNA)
pRGEB31
gRNA_AAVS1-T2
pCFD4-U6-1_U6-3tandemgRNAs
pDR274
pRGEB32
gRNA_Cloning Vector
pCLIP-All-EFS-Ametrine
Peft-3_cas9-SV40_NLS_tbb-2 3'UTR
pROSA26-Puro-DNR
gRNA_GFP-T1
pCLIP-All-EFS-Blast
pET-28b-Cas9-His
pRPR1_gRNA_handle_RPR1t
gRNA_GFP-T2
pCLIP-All-EFS-Puro
pET-Cas9-6xHis
pSECC
HBT-pcoCas9
pCLIP-All-EFS-tRFP
pET-Cas9-NLS-6xHis
pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s)
hCas9
pCLIP-All-EFS-ZsGreen
pET-NLS-Cas9-6xHis
pSLQ1658-dCas9-EGFP
hCas9_D10A
pCLIP-All-hCMV-Blast
pET28a Cas9-Cys
pSpCas9(BB)-2A-GFP (PX458)
JDS246
pCLIP-All-hCMV-Puro
pFGC-pcoCas9
pSpCas9(BB)-2A-Puro (PX459) V2.0
lenti dCAS-VP64_Blast
pCLIP-All-hCMV-tRFP
pGL3-U6-sgRNA-PGK-puromycin
pSpCas9n(BB)-2A-GFP (PX461)
lenti MS2-P65-HSF1_Hygro
pCLIP-All-hCMV-ZsGreen
pgRNA-bacteria
pSpCas9n(BB)-2A-Puro (PX462) V2.0
lenti sgRNA(MS2)_zeo backbone
pCLIP-All-Nickase(D10A)-EFS-Blast
pgRNA-humanized
pSpCas9n(BB) (PX460)
lentiCas9-Blast
pCLIP-All-Nickase(D10A)-EFS-Puro
pGuide
pSpCas9 (PX165)
lentiCas9-EGFP
pCLIP-All-Nickase(D10A)-EFS-tRFP
pGuide-it-sgRNA1 Vector (Linear)
pSPgRNA
lentiCas9n(D10A)-Blast
pCLIP-All-Nickase(D10A)-EFS-ZsGreen
pGuide-it-tdTomato Vector (Linear)
pSQT834
lentiCRISPR v2
pCLIP-All-Nickase(D10A)-hCMV-Puro
pGuide-it-ZsGreen1 Vector (Linear)
pSQT1313
lentiGuide-Puro
pCLIP-All-Nickase(D10A)-hCMV-tRFP
pHAGE EF1-alpha dCas9-KRAB
pST1374-N-NLS-flag-linker-Cas9-D10A
LSL-Cas9-Rosa26TV
pCLIP-All-Nickase(D10A)-hCMV-ZsGreen
pHAGE EF1-alpha dCas9-VP64
pST1374-N-NLS-flag-linker-Cas9-H840A
M-SPcas
pCLIP-All-Nickase(D10A)-TRE3G-Blast
pHAGE TRE dCas9
pST1374-NLS-flag-linker-Cas9
M-ST1cas
pCLIP-All-Nickase(D10A)-TRE3G-Puro
pHAGE TRE dCas9-KRAB
pT3TS-nCas9n
MLM3613
pCLIP-All-Nickase(D10A)-TRE3G-tRFP
pHAGE TRE dCas9-VP64
pT7-Cas9
MLM3636
pCLIP-All-Nickase(D10A)-TRE3G-ZsGreen
pHAGE-TO-dCas9-3XGFP
pT7-Cas9-Nickase
MLM3639
pCLIP-All-TRE3G-Blast
pHAGE-TO-dCas9-3XmCherry
pT7-gRNA
MS2-P65-HSF1_GFP
pCLIP-All-TRE3G-Puro
pHAGE-TO-nls-st1dCas9-3nls-3XTagBFP2
pT7-Guide-IVT
MSP1101
pCLIP-All-TRE3G-tRFP
pHAGE-TO-nmdCas9-3XGFP
pTDH3-dCas9
MSP2133
pCLIP-All-TRE3G-ZsGreen
pHAGE-TO-nmdCas9-3XmCherry
pTDH3-dCas9-Mxi1
MSP2135
pCLIP-Cas9(D10a)-Nickase-EFS-Puro
pHR-SFFV-dCas9-BFP
pTOL-hCMV-TET3G-Hygro Transactivator
MSP469
pCLIP-Cas9(D10a)-Nickase-EFS-Blast
pHR-SFFV-dCas9-BFP-KRAB
pTPGI_dCas9_VP64
p3s-Cas9HC
pCLIP-Cas9(D10a)-Nickase-EFS-tRFP
pHR-SFFV-KRAB-dCas9-P2A-mCherry
pU6-BbsI-chiRNA
p414-TEF1p-Cas9-CYC1t
pCLIP-Cas9(D10a)-Nickase-EFS-ZsGreen
pHRdSV40-dCas9-10xGCN4_v4-P2A-BFP
PU6_klp-12_sgRNA
p415-GalL-Cas9-CYC1t
pCLIP-Cas9(D10a)-Nickase-hCMV-Blast