pHR-SFFV-KRAB-dCas9-P2A-mCherry

Lentiviral CRISPR interference vector for expressing mCherry together with a KRAB domain fused to catalytically inactive dCas9.

Sequence Author: Weissman Lab / Addgene #60954

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HomePlasmidsCRISPR PlasmidspHR-SFFV-KRAB-dCas9-P2A-mCherry
SV40 NLS SV40 NLS ATG cPPT/CTS pHR-SFFV-KRAB-dCas9-P2A-mCherry 14,247 bp
ATG
8354 .. 8356  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
8354 .. 8356  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
KRAB
8384 .. 8569  =  186 bp
62 amino acids  =  7.4 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
KRAB
8384 .. 8569  =  186 bp
62 amino acids  =  7.4 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
dCas9
8591 .. 12,694  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   8591 .. 8617  =  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
8591 .. 12,694  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   8618 .. 8620  =  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
8591 .. 12,694  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   8621 .. 11,107  =  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
8591 .. 12,694  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   11,108 .. 11,110  =  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
8591 .. 12,694  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   11,111 .. 12,694  =  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
8591 .. 12,694  =  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
HA
12,698 .. 12,724  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
12,698 .. 12,724  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
SV40 NLS
12,743 .. 12,763  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
12,743 .. 12,763  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
12,770 .. 12,790  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
12,770 .. 12,790  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
P2A
12,830 .. 12,886  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
P2A
12,830 .. 12,886  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
mCherry
12,887 .. 13,597  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
12,887 .. 13,597  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
AmpR
1277 .. 2137  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1277 .. 1345  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1277 .. 2137  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1346 .. 2137  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1277 .. 2137  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
LTR
5548 .. 6181  =  634 bp
long terminal repeat (LTR) from HIV-1
LTR
5548 .. 6181  =  634 bp
long terminal repeat (LTR) from HIV-1
ori
2308 .. 2896  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2308 .. 2896  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
WPRE
13,657 .. 14,245  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
13,657 .. 14,245  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
SFFV promoter
7866 .. 8273  =  408 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
SFFV promoter
7866 .. 8273  =  408 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
SV40 promoter
3142 .. 3471  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3142 .. 3471  =  330 bp
SV40 enhancer and early promoter
LTR (ΔU3)
108 .. 341  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
LTR (ΔU3)
108 .. 341  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
RRE
6850 .. 7082  =  233 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
6850 .. 7082  =  233 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
SV40 poly(A) signal
5245 .. 5379  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5245 .. 5379  =  135 bp
SV40 polyadenylation signal
HIV-1 Ψ
6228 .. 6353  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
6228 .. 6353  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
7596 .. 7713  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
7596 .. 7713  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
1172 .. 1276  =  105 bp
AmpR promoter
1172 .. 1276  =  105 bp
SV40 ori
3322 .. 3457  =  136 bp
SV40 origin of replication
SV40 ori
3322 .. 3457  =  136 bp
SV40 origin of replication
ORF:  574 .. 921  =  348 bp
ORF:  115 amino acids  =  11.9 kDa
ORF:  3754 .. 4128  =  375 bp
ORF:  124 amino acids  =  14.0 kDa
ORF:  7189 .. 7788  =  600 bp
ORF:  199 amino acids  =  23.6 kDa
ORF:  13,747 .. 27  =  528 bp
ORF:  175 amino acids  =  19.3 kDa
ORF:  1277 .. 2137  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3908 .. 4189  =  282 bp
ORF:  93 amino acids  =  10.7 kDa
ORF:  4220 .. 4603  =  384 bp
ORF:  127 amino acids  =  14.9 kDa
ORF:  6728 .. 7066  =  339 bp
ORF:  112 amino acids  =  12.4 kDa
ORF:  7886 .. 8137  =  252 bp
ORF:  83 amino acids  =  9.2 kDa
ORF:  8354 .. 13,597  =  5244 bp
ORF:  1747 amino acids  =  200.6 kDa
ORF:  13,760 .. 187  =  675 bp
ORF:  224 amino acids  =  24.0 kDa
ORF:  4749 .. 5222  =  474 bp
ORF:  157 amino acids  =  18.2 kDa
ORF:  490 .. 801  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  1741 .. 2007  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3802 .. 4221  =  420 bp
ORF:  139 amino acids  =  15.9 kDa
ORF:  6562 .. 7023  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  9940 .. 10,233  =  294 bp
ORF:  97 amino acids  =  10.7 kDa
ORF:  10,504 .. 10,953  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  12,718 .. 12,954  =  237 bp
ORF:  78 amino acids  =  8.5 kDa
ORF:  7694 .. 8101  =  408 bp
ORF:  135 amino acids  =  15.2 kDa
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pDCC6
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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
pHRdSV40-NLS-dCas9-24xGCN4_v4-NLS-P2A-BFP-dWPRE
pU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E1B
p426-SNR52p-gRNA.CAN1.Y-SUP4t
pCLIP-Cas9(D10a)-Nickase-hCMV-Puro
pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS
pU6-(BbsI)_CBh-Cas9-T2A-mcherry-P2A-Ad4E4orf6
pAAV-Guide-it-1 Vector (Linear)
pCLIP-Cas9(D10a)-Nickase-hCMV-tRFP
pHSN401
pUC119-gRNA
pAAV-Guide-it-Down Vector (Linear)
pCLIP-Cas9(D10a)-Nickase-hCMV-ZsGreen
pHsp70-Cas9
pUC57-sgRNA expression vector
pAAV-Guide-it-Up Vector
pCLIP-Cas9(D10a)-Nickase-TRE3G-Blast
pICH41308_hCas9
Puro-Cas9 donor
pAAVS1-BSD-DNR
pCLIP-Cas9(D10a)-Nickase-TRE3G-Puro
pICH47742_2x35S-5'UTR-hCas9(STOP)-NOST
pWJ66
pAAVS1-Puro-DNR
pCLIP-Cas9(D10a)-Nickase-TRE3G-tRFP
piCRg Entry
pWJ68
pAC2-dual-dCas9VP48-sgExpression
pCLIP-Cas9(D10a)-Nickase-TRE3G-ZsGreen
pICSL01009_AtU6p
pwtCas9-bacteria
pAc-sgRNA-Cas9
pCLIP-Cas9-Nuclease-EFS-Blast
pK7WGF2_hCas9
pX260
pAC149-pCR8-dCas9VP160
pCLIP-Cas9-Nuclease-EFS-Puro
pKLV-U6gRNA(BbsI)-PGKpuro2ABFP
pX330
pAC152-dual-dCas9VP64-sgExpression
pCLIP-Cas9-Nuclease-EFS-tRFP
pL-CRISPR.EFS.GFP
pX330A-1x2
pAC153-dual-dCas9VP96-sgExpression
pCLIP-Cas9-Nuclease-EFS-ZsGreen
pL-CRISPR.EFS.tRFP
pX330S-2
pAC154-dual-dCas9VP160-sgExpression
pCLIP-Cas9-Nuclease-hCMV-Blast
pL-CRISPR.SFFV.GFP
pX334
pAC155-pCR8-sgExpression
pCLIP-Cas9-Nuclease-hCMV-Puro
pL-CRISPR.SFFV.tRFP
pX335
pAC84-pCR8-dCas9
pCLIP-Cas9-Nuclease-hCMV-tRFP
pLenti-Cas9
PX551
pAC91-pmax-dCas9VP64
pCLIP-Cas9-Nuclease-hCMV-ZsGreen
pLenti-Cas9-IRES-Puro
PX552
pAC93-pmax-dCas9VP160
pCLIP-Cas9-Nuclease-TRE3G-Blast
pLenti-Cas9-P2A-tGFP
pX600
pAC94-pmax-dCas9VP160-2A-puro
pCLIP-Cas9-Nuclease-TRE3G-Puro
pLenti-Cas-Guide
pX601
pAC95-pmax-dCas9VP160-2A-neo
pCLIP-Cas9-Nuclease-TRE3G-tRFP
pLenti-EF1a-Cas9-IRES-Puro
pX602
pACT2-CAS9c
pCLIP-Cas9-Nuclease-TRE3G-ZsGreen
pLenti-Guide-Puro
PX851
pBS-Hsp70-Cas9
pCLIP-gRNA-EFS-Blast
pLKO.1-puro U6 sgRNA BfuAI stuffer
PX852
pCAG-T3-hCAS-pA
pCLIP-gRNA-EFS-Puro
pLSC-5
PX853
pCAG-T3-hCasD10A-pA
pCLIP-gRNA-EFS-tRFP
pLV hUbC-Cas9-T2A-GFP
PX854
pCas
pCLIP-gRNA-EFS-ZsGreen
pLV hUbC-dCas9 VP64-T2A-GFP
PX855
pCas-Guide-AAVS1
pCLIP-gRNA-hCMV-Blast
pLV hUbC-dCas9-T2A-GFP
PX856
pCas-Guide-CRISPRa
pCLIP-gRNA-hCMV-Puro
pLVX-hyg-sgRNA1 Vector (Linear)
pY010 (pcDNA3.1-hAsCpf1)
pCas-Guide-CRISPRi
pCLIP-gRNA-hCMV-tRFP
pLVX-puro-Cas9
pY016 (pcDNA3.1-hLbCpf1)
pCas-Guide-EF1a-CD4
pCLIP-gRNA-hCMV-ZsGreen
pLX-sgRNA
sgRNA(MS2) cloning backbone
pCas-Guide-EF1a-GFP
pCRCT
pMB66
SP-Cas9
pCas-Guide-Nickase
pCRISPomyces-1
pMB70
SP-dCas9-VPR
pCas-Guide-ROSA26
pCRISPomyces-2
pME-Cas9
VP12
pCas9
pCRISPR

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