pRGEB31

Plant binary vector for stably expressing a single guide RNA (sgRNA) together with Cas9.

Sequence Author: Yang Lab / Addgene #51295

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BsaI (382) BsaI (402) polIII terminator attB1 ATG 3xFLAG SV40 NLS RB T-DNA repeat pRGEB31 15,037 bp
BsaI  (382)
2 sites		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BsaI  (402)
2 sites		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
ATG
1477 .. 1479  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1477 .. 1479  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
1480 .. 1545  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
1480 .. 1545  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
1552 .. 1572  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1552 .. 1572  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
1597 .. 5697  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
1597 .. 5697  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
nucleoplasmin NLS
5698 .. 5745  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
5698 .. 5745  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
pVS1 RepA
8742 .. 9809  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 RepA
8742 .. 9809  =  1068 bp
355 amino acids  =  39.7 kDa
Product: replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
HygR
12,949 .. 13,974  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
12,949 .. 13,974  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
KanR
11,414 .. 12,208  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
11,414 .. 12,208  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
14,042 .. 14,719  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
14,042 .. 14,719  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
pVS1 StaA
7679 .. 8308  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 StaA
7679 .. 8308  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
ori
10,739 .. 11,327  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
10,739 .. 11,327  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
OsU3 promoter
6 .. 386  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
OsU3 promoter
6 .. 386  =  381 bp
Oryza sativa (rice) snRNA U3 promoter
CaMV 35S promoter
1044 .. 1389  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
1044 .. 1389  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
NOS terminator
5858 .. 6110  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
5858 .. 6110  =  253 bp
nopaline synthase terminator and poly(A) signal
pVS1 oriV
9875 .. 10,069  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 oriV
9875 .. 10,069  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
12,735 .. 12,909  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
12,735 .. 12,909  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
10,413 .. 10,553  =  141 bp
basis of mobility region from pBR322
bom
10,413 .. 10,553  =  141 bp
basis of mobility region from pBR322
gRNA scaffold
403 .. 478  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
403 .. 478  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
attB1
1411 .. 1435  =  25 bp
recombination site for the Gateway® BP reaction
attB1
1411 .. 1435  =  25 bp
recombination site for the Gateway® BP reaction
attB2
5778 .. 5802  =  25 bp
recombination site for the Gateway® BP reaction
attB2
5778 .. 5802  =  25 bp
recombination site for the Gateway® BP reaction
RB T-DNA repeat
6355 .. 6379  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
6355 .. 6379  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
12,633 .. 12,657  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
12,633 .. 12,657  =  25 bp
left border repeat from nopaline C58 T-DNA
polIII terminator
479 .. 484  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
479 .. 484  =  6 bp
RNA polymerase III transcription terminator
ORF:  541 .. 1290  =  750 bp
ORF:  249 amino acids  =  28.0 kDa
ORF:  1477 .. 5748  =  4272 bp
ORF:  1423 amino acids  =  164.6 kDa
ORF:  6694 .. 7380  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  12,901 .. 13,641  =  741 bp
ORF:  246 amino acids  =  27.0 kDa
ORF:  584 .. 814  =  231 bp
ORF:  76 amino acids  =  9.1 kDa
ORF:  7679 .. 8308  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  8703 .. 9809  =  1107 bp
ORF:  368 amino acids  =  41.2 kDa
ORF:  11,547 .. 11,777  =  231 bp
ORF:  76 amino acids  =  8.3 kDa
ORF:  7145 .. 7417  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  8576 .. 8839  =  264 bp
ORF:  87 amino acids  =  9.1 kDa
ORF:  11,414 .. 12,208  =  795 bp
ORF:  264 amino acids  =  31.0 kDa
ORF:  4 .. 342  =  339 bp
ORF:  112 amino acids  =  12.7 kDa
ORF:  1471 .. 4113  =  2643 bp
ORF:  880 amino acids  =  92.7 kDa
ORF:  7417 .. 7731  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  12,949 .. 13,974  =  1026 bp
ORF:  341 amino acids  =  38.0 kDa
ORF:  14,146 .. 14,463  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  14,473 .. 14,931  =  459 bp
ORF:  152 amino acids  =  16.9 kDa
ORF:  2943 .. 3236  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  3507 .. 3956  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  6726 .. 7076  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  8583 .. 9176  =  594 bp
ORF:  197 amino acids  =  22.2 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-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
pCLIP-gRNA-hCMV-Blast