pLVX-puro-Cas9

Lentiviral Cas9 expression vector with the constitutive CMV promoter and a puromycin resistance marker.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsCRISPR PlasmidspLVX-puro-Cas9
SgrDI (11,937) AhdI (11,016) KpnI (8956) Acc65I (8952) CsiI - SexAI * (8181) BstEII (7748) RsrII (7730) BsiWI (7670) SpeI (7339) AgeI (7204) SphI (7133) XbaI (7078) AscI (6893) EcoRI (6760) PaqCI (6198) FseI (1150) MfeI (1189) BbvCI (1424) AleI (1577) cPPT/CTS BspDI - ClaI (2180) SnaBI (2480) ATG SV40 NLS SbfI (3185) TspMI - XmaI (4304) SmaI (4306) BamHI (4943) BstXI (5727) FspAI (5888) pLVX-puro-Cas9 12,305 bp
SgrDI  (11,937)
1 site		 C G T C G A C G G C A G C T G C
AhdI  (11,016)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
KpnI  (8956)
1 site		 G G T A C C C C A T G G
Acc65I  (8952)
1 site		 G G T A C C C C A T G G
CsiI  (8181)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (8181)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BstEII  (7748)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
RsrII  (7730)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsiWI  (7670)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
SpeI  (7339)
1 site		 A C T A G T T G A T C A
AgeI  (7204)
1 site		 A C C G G T T G G C C A
SphI  (7133)
1 site		 G C A T G C C G T A C G
XbaI  (7078)
1 site		 T C T A G A A G A T C T
AscI  (6893)
1 site		 G G C G C G C C C C G C G C G G
EcoRI  (6760)
1 site		 G A A T T C C T T A A G
PaqCI  (6198)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
FseI  (1150)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
MfeI  (1189)
1 site		 C A A T T G G T T A A C
BbvCI  (1424)
1 site		 C C T C A G C G G A G T C G
AleI  (1577)
1 site		 C A C N N N N G T G G T G N N N N C A C
BspDI  (2180)
1 site		 A T C G A T T A G C T A
ClaI  (2180)
1 site		 A T C G A T T A G C T A
SnaBI  (2480)
1 site		 T A C G T A A T G C A T
SbfI  (3185)
1 site		 C C T G C A G G G G A C G T C C
TspMI  (4304)
1 site		 C C C G G G G G G C C C
XmaI  (4304)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (4306)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BamHI  (4943)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BstXI  (5727)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
FspAI  (5888)
1 site		 R T G C G C A Y Y A C G C G T R
ATG
2821 .. 2823  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2821 .. 2823  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
2824 .. 2889  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
2824 .. 2889  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
2896 .. 2916  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2896 .. 2916  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
2941 .. 7041  =  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
2941 .. 7041  =  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
SV40 NLS
7054 .. 7074  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
7054 .. 7074  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
AmpR
10,943 .. 11,803  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   10,943 .. 11,734  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,943 .. 11,803  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   11,735 .. 11,803  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,943 .. 11,803  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
3' LTR
9023 .. 9656  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
9023 .. 9656  =  634 bp
3' long terminal repeat (LTR) from HIV-1
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
PuroR
7614 .. 8213  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
7614 .. 8213  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
8227 .. 8815  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
8227 .. 8815  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
10,187 .. 10,772  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
10,187 .. 10,772  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
7094 .. 7593  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
7094 .. 7593  =  500 bp
mouse phosphoglycerate kinase 1 promoter
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2201 .. 2504  =  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.
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2505 .. 2708  =  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
2028 .. 2143  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2143  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
11,804 .. 11,908  =  105 bp
AmpR promoter
11,804 .. 11,908  =  105 bp
ORF:  2821 .. 7077  =  4257 bp
ORF:  1418 amino acids  =  164.2 kDa
ORF:  8317 .. 8844  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  8330 .. 8626  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  7374 .. 8213  =  840 bp
ORF:  279 amino acids  =  30.1 kDa
ORF:  11,073 .. 11,339  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  10,943 .. 11,803  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  2809 .. 3417  =  609 bp
ORF:  202 amino acids  =  22.5 kDa
ORF:  4342 .. 4860  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  4924 .. 5457  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
ORF:  6451 .. 6984  =  534 bp
ORF:  177 amino acids  =  20.0 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
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

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
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TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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