pHAGE EF1-alpha dCas9-KRAB

Lentiviral CRISPR interference vector for constitutive expression of catalytically inactive dCas9 fused to a KRAB transcriptional repressor domain.

Sequence Author: Maehr

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HomePlasmidsCRISPR PlasmidspHAGE EF1-alpha dCas9-KRAB
pHAGE EF1 α dCas9-KRAB 12,843 bp
dCas9
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   8335 .. 8361  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   8362 .. 8364  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   8365 .. 10,851  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   10,852 .. 10,854  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   10,855 .. 12,438  =  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
8335 .. 12,438  =  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
12,451 .. 12,471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
12,451 .. 12,471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
3xHA
12,472 .. 12,561  =  90 bp
30 amino acids  =  3.5 kDa
Product: three tandem HA epitope tags
3xHA
12,472 .. 12,561  =  90 bp
30 amino acids  =  3.5 kDa
Product: three tandem HA epitope tags
KRAB
12,598 .. 12,792  =  195 bp
65 amino acids  =  7.8 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
KRAB
12,598 .. 12,792  =  195 bp
65 amino acids  =  7.8 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
EF-1α promoter
7088 .. 8266  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
7088 .. 8266  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
2464 .. 3324  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   2464 .. 2532  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2464 .. 3324  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2533 .. 3324  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2464 .. 3324  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
LTR
4869 .. 5502  =  634 bp
long terminal repeat (LTR) from HIV-1
LTR
4869 .. 5502  =  634 bp
long terminal repeat (LTR) from HIV-1
PuroR
646 .. 1245  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
646 .. 1245  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
1258 .. 1846  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
1258 .. 1846  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
3495 .. 4083  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3495 .. 4083  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
126 .. 625  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
126 .. 625  =  500 bp
mouse phosphoglycerate kinase 1 promoter
LTR (ΔU3)
1921 .. 2154  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
LTR (ΔU3)
1921 .. 2154  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
RRE
6171 .. 6404  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
6171 .. 6404  =  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
5549 .. 5674  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
5549 .. 5674  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
6926 .. 7043  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
6926 .. 7043  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
2359 .. 2463  =  105 bp
AmpR promoter
2359 .. 2463  =  105 bp
HA
67 .. 93  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
67 .. 93  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
attB1
8301 .. 8325  =  25 bp
recombination site for the Gateway® BP reaction
attB1
8301 .. 8325  =  25 bp
recombination site for the Gateway® BP reaction
attB2
12,833 .. 14  =  25 bp
recombination site for the Gateway® BP reaction
attB2
12,833 .. 14  =  25 bp
recombination site for the Gateway® BP reaction
EF-1α intron A
7319 .. 8257  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
7319 .. 8257  =  939 bp
intron upstream of the start codon of human EF-1α
ORF:  406 .. 1245  =  840 bp
ORF:  279 amino acids  =  30.1 kDa
ORF:  1348 .. 1860  =  513 bp
ORF:  170 amino acids  =  18.6 kDa
ORF:  2464 .. 3324  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  6049 .. 6975  =  927 bp
ORF:  308 amino acids  =  35.4 kDa
ORF:  7723 .. 7956  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  8335 .. 12,795  =  4461 bp
ORF:  1486 amino acids  =  171.9 kDa
ORF:  1361 .. 1894  =  534 bp
ORF:  177 amino acids  =  18.8 kDa
ORF:  12,401 .. 12,739  =  339 bp
ORF:  112 amino acids  =  13.2 kDa
ORF:  6906 .. 7196  =  291 bp
ORF:  96 amino acids  =  10.5 kDa
ORF:  7644 .. 8081  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  8254 .. 8745  =  492 bp
ORF:  163 amino acids  =  17.9 kDa
ORF:  9739 .. 10,257  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  10,321 .. 10,596  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  11,224 .. 12,381  =  1158 bp
ORF:  385 amino acids  =  43.7 kDa
ORF:  2928 .. 3194  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5883 .. 6344  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  7386 .. 8006  =  621 bp
ORF:  206 amino acids  =  23.0 kDa
ORF:  6749 .. 6997  =  249 bp
ORF:  82 amino acids  =  9.0 kDa
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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)
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pHsp70-Cas9
pUC57-sgRNA expression vector
pAAV-Guide-it-Up Vector
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pICH41308_hCas9
Puro-Cas9 donor
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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
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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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