pHAGE-TO-nls-st1dCas9-3nls-3XTagBFP2

Lentiviral vector for labeling chromosomal loci by tetracycline-regulatable expression of catalytically dead Streptococcus thermophilus dCas9 fused to three copies of mTagBFP2.

Sequence Author: Pederson Lab / Addgene #64512

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HomePlasmidsCRISPR PlasmidspHAGE-TO-nls-st1dCas9-3nls-3XTagBFP2
SV40 NLS SV40 NLS CMV promoter tet operator tet operator ATG SV40 NLS pHAGE-TO-nls-st1dCas9-3nls-3XTagBFP2 12,284 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
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
   Segment 1:  
   3016 .. 3036  =  21 bp
   7 amino acids  =  715.8 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
   Segment 2:  
   3037 .. 3039  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
   Segment 3:  
   3040 .. 4803  =  1764 bp
   588 amino acids  =  67.5 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
   Segment 4:  
   4804 .. 4809  =  6 bp
   2 amino acids  =  160.2 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
   Segment 5:  
   4810 .. 4875  =  66 bp
   22 amino acids  =  2.4 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
   Segment 6:  
   4876 .. 4878  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
   Segment 7:  
   4879 .. 6375  =  1497 bp
   499 amino acids  =  58.3 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
St1dCas9
3016 .. 6375  =  3360 bp
1120 amino acids  =  129.1 kDa
7 segments
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D9A, D598A, H599A, and N622A
SV40 NLS
6391 .. 6411  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6391 .. 6411  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6424 .. 6444  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6424 .. 6444  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6451 .. 6471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6451 .. 6471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
mTagBFP2
6484 .. 7194  =  711 bp
237 amino acids  =  26.7 kDa
Product: enhanced monomeric blue fluorescent protein (Subach et al., 2011)
mammalian codon-optimized
mTagBFP2
6484 .. 7194  =  711 bp
237 amino acids  =  26.7 kDa
Product: enhanced monomeric blue fluorescent protein (Subach et al., 2011)
mammalian codon-optimized
mTagBFP2
7213 .. 7923  =  711 bp
237 amino acids  =  26.7 kDa
Product: enhanced monomeric blue fluorescent protein (Subach et al., 2011)
mammalian codon-optimized
mTagBFP2
7213 .. 7923  =  711 bp
237 amino acids  =  26.7 kDa
Product: enhanced monomeric blue fluorescent protein (Subach et al., 2011)
mammalian codon-optimized
mTagBFP2
7942 .. 8652  =  711 bp
237 amino acids  =  26.7 kDa
Product: enhanced monomeric blue fluorescent protein (Subach et al., 2011)
mammalian codon-optimized
mTagBFP2
7942 .. 8652  =  711 bp
237 amino acids  =  26.7 kDa
Product: enhanced monomeric blue fluorescent protein (Subach et al., 2011)
mammalian codon-optimized
AmpR
9880 .. 10,740  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   9880 .. 9948  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9880 .. 10,740  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   9949 .. 10,740  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9880 .. 10,740  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
8674 .. 9262  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
8674 .. 9262  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
10,911 .. 11,499  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
10,911 .. 11,499  =  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.
3' LTR (ΔU3)
9337 .. 9570  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
9337 .. 9570  =  234 bp
self-inactivating 3' 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
9775 .. 9879  =  105 bp
AmpR promoter
9775 .. 9879  =  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 .. 8661  =  5682 bp
ORF:  1893 amino acids  =  215.4 kDa
ORF:  8764 .. 9276  =  513 bp
ORF:  170 amino acids  =  18.6 kDa
ORF:  9880 .. 10,740  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1181 .. 2107  =  927 bp
ORF:  308 amino acids  =  35.4 kDa
ORF:  2762 .. 3073  =  312 bp
ORF:  103 amino acids  =  12.3 kDa
ORF:  8777 .. 9310  =  534 bp
ORF:  177 amino acids  =  18.8 kDa
ORF:  1881 .. 2129  =  249 bp
ORF:  82 amino acids  =  9.0 kDa
ORF:  4140 .. 4415  =  276 bp
ORF:  91 amino acids  =  10.2 kDa
ORF:  4443 .. 4826  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  6300 .. 6539  =  240 bp
ORF:  79 amino acids  =  9.0 kDa
ORF:  10,344 .. 10,610  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  6448 .. 8460  =  2013 bp
ORF:  670 amino acids  =  69.7 kDa
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Individual Sequences & Maps

3xFLAG-dCas9 pCMV-7.1
pCas9
pCRISPRa-Enhancer
pME-Cas9-T2A-GFP
3xFLAG-dCas9 pMXs-neo
pCas9_GFP
pCS2+hSpCas9
pMJ806
3xFLAG-dCas9 pMXs-puro
pCas9D10A_GFP
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 (PX165)
JDS246
pCLIP-All-hCMV-Puro
pFGC-pcoCas9
pSpCas9(BB)-2A-GFP (PX458)
lenti dCAS-VP64_Blast
pCLIP-All-hCMV-tRFP
pGL3-U6-sgRNA-PGK-puromycin
pSpCas9(BB)-2A-Puro (PX459) V2.0
lenti MS2-P65-HSF1_Hygro
pCLIP-All-hCMV-ZsGreen
pgRNA-bacteria
pSpCas9n(BB) (PX460)
lenti sgRNA(MS2)_zeo backbone
pCLIP-All-Nickase(D10A)-EFS-Blast
pgRNA-humanized
pSpCas9n(BB)-2A-GFP (PX461)
lentiCas9-Blast
pCLIP-All-Nickase(D10A)-EFS-Puro
pGuide
pSpCas9n(BB)-2A-Puro (PX462) V2.0
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)
pSQT1313
lentiCRISPR v2
pCLIP-All-Nickase(D10A)-hCMV-Puro
pGuide-it-ZsGreen1 Vector (Linear)
pSQT834
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)_CBh-Cas9-T2A-BFP-P2A-Ad4E1B
p414-TEF1p-Cas9-CYC1t
pCLIP-Cas9(D10a)-Nickase-EFS-ZsGreen
pHRdSV40-dCas9-10xGCN4_v4-P2A-BFP
pU6-(BbsI)_CBh-Cas9-T2A-mcherry-P2A-Ad4E4orf6
p415-GalL-Cas9-CYC1t
pCLIP-Cas9(D10a)-Nickase-hCMV-Blast
pHRdSV40-NLS-dCas9-24xGCN4_v4-NLS-P2A-BFP-dWPRE
pU6-BbsI-chiRNA
p426-SNR52p-gRNA.CAN1.Y-SUP4t
pCLIP-Cas9(D10a)-Nickase-hCMV-Puro
pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS
PU6_klp-12_sgRNA
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
pAc-sgRNA-Cas9
pCLIP-Cas9(D10a)-Nickase-TRE3G-ZsGreen
pICSL01009_AtU6p
pwtCas9-bacteria
pAC149-pCR8-dCas9VP160
pCLIP-Cas9-Nuclease-EFS-Blast
pK7WGF2_hCas9
pX260
pAC152-dual-dCas9VP64-sgExpression
pCLIP-Cas9-Nuclease-EFS-Puro
pKLV-U6gRNA(BbsI)-PGKpuro2ABFP
pX330
pAC153-dual-dCas9VP96-sgExpression
pCLIP-Cas9-Nuclease-EFS-tRFP
pL-CRISPR.EFS.GFP
pX330A-1x2
pAC154-dual-dCas9VP160-sgExpression
pCLIP-Cas9-Nuclease-EFS-ZsGreen
pL-CRISPR.EFS.tRFP
pX330S-2
pAC155-pCR8-sgExpression
pCLIP-Cas9-Nuclease-hCMV-Blast
pL-CRISPR.SFFV.GFP
pX334
pAC2-dual-dCas9VP48-sgExpression
pCLIP-Cas9-Nuclease-hCMV-Puro
pL-CRISPR.SFFV.tRFP
pX335
pAC84-pCR8-dCas9
pCLIP-Cas9-Nuclease-hCMV-tRFP
pLenti-Cas-Guide
PX551
pAC91-pmax-dCas9VP64
pCLIP-Cas9-Nuclease-hCMV-ZsGreen
pLenti-Cas9
PX552
pAC93-pmax-dCas9VP160
pCLIP-Cas9-Nuclease-TRE3G-Blast
pLenti-Cas9-IRES-Puro
pX600
pAC94-pmax-dCas9VP160-2A-puro
pCLIP-Cas9-Nuclease-TRE3G-Puro
pLenti-Cas9-P2A-tGFP
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-