pL-CRISPR.SFFV.tRFP

Lentiviral CRISPR/Cas9 vector with a TagRFP marker driven by the SFFV promoter, for genome editing in mammalian cells.

Sequence Author: Ebert Lab / Addgene #57826

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HomePlasmidsCRISPR PlasmidspL-CRISPR.SFFV.tRFP
BsmBI (1961) BsmBI (3846) polIII terminator ATG FLAG SV40 NLS pL-CRISPR.SFFV.tRFP 13,729 bp
BsmBI  (1961)
2 sites		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
BsmBI  (3846)
2 sites		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
ATG
4558 .. 4560  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4558 .. 4560  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
FLAG
4561 .. 4584  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
4561 .. 4584  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
SV40 NLS
4591 .. 4611  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4591 .. 4611  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
4636 .. 8736  =  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
4636 .. 8736  =  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
8737 .. 8784  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
8737 .. 8784  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
P2A
8800 .. 8856  =  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
8800 .. 8856  =  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.
TagRFP
8857 .. 9570  =  714 bp
237 amino acids  =  26.7 kDa
Product: monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)
mammalian codon-optimized
TagRFP
8857 .. 9570  =  714 bp
237 amino acids  =  26.7 kDa
Product: monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)
mammalian codon-optimized
spacer
1962 .. 3846  =  1885 bp
spacer
1962 .. 3846  =  1885 bp
AmpR
11,646 .. 12,506  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   11,646 .. 11,714  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
11,646 .. 12,506  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   11,715 .. 12,506  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
11,646 .. 12,506  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
12,677 .. 13,265  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
12,677 .. 13,265  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
WPRE
9586 .. 10,174  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
9586 .. 10,174  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
f1 ori
11,060 .. 11,515  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
11,060 .. 11,515  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SFFV promoter
4145 .. 4546  =  402 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
SFFV promoter
4145 .. 4546  =  402 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
U6 promoter
1717 .. 1957  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
1717 .. 1957  =  241 bp
RNA polymerase III promoter for human U6 snRNA
3' LTR (ΔU3)
10,246 .. 10,479  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
10,246 .. 10,479  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
RRE
1077 .. 1310  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1077 .. 1310  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RSV promoter
5 .. 231  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
5 .. 231  =  227 bp
Rous sarcoma virus enhancer/promoter
5' LTR (truncated)
232 .. 412  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
232 .. 412  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
10,718 .. 10,853  =  136 bp
SV40 origin of replication
SV40 ori
10,718 .. 10,853  =  136 bp
SV40 origin of replication
HIV-1 Ψ
459 .. 584  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
459 .. 584  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
10,557 .. 10,678  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
10,557 .. 10,678  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
3970 .. 4087  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
3970 .. 4087  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
11,541 .. 11,645  =  105 bp
AmpR promoter
11,541 .. 11,645  =  105 bp
gRNA scaffold
3847 .. 3922  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
3847 .. 3922  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
polIII terminator
3923 .. 3928  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3923 .. 3928  =  6 bp
RNA polymerase III transcription terminator
ORF:  955 .. 1830  =  876 bp
ORF:  291 amino acids  =  33.6 kDa
ORF:  4165 .. 4416  =  252 bp
ORF:  83 amino acids  =  9.2 kDa
ORF:  4558 .. 9570  =  5013 bp
ORF:  1670 amino acids  =  192.0 kDa
ORF:  9676 .. 10,185  =  510 bp
ORF:  169 amino acids  =  18.5 kDa
ORF:  10,579 .. 10,815  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  13,663 .. 179  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  3950 .. 4219  =  270 bp
ORF:  89 amino acids  =  10.4 kDa
ORF:  9689 .. 10,219  =  531 bp
ORF:  176 amino acids  =  18.7 kDa
ORF:  11,646 .. 12,506  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  13,590 .. 160  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  12,110 .. 12,376  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2140 .. 2535  =  396 bp
ORF:  131 amino acids  =  14.0 kDa
ORF:  4096 .. 4380  =  285 bp
ORF:  94 amino acids  =  10.3 kDa
ORF:  4501 .. 7152  =  2652 bp
ORF:  883 amino acids  =  92.8 kDa
ORF:  870 .. 1250  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  5982 .. 6275  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  6546 .. 6995  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  8970 .. 9215  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
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Individual Sequences & Maps

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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
pLV hUbC-dCas9-T2A-GFP
PX856
pCas-Guide-AAVS1
pCLIP-gRNA-hCMV-Puro
pLVX-hyg-sgRNA1 Vector (Linear)
pY010 (pcDNA3.1-hAsCpf1)
pCas-Guide-CRISPRa