pAAVS1-Puro-DNR

Mammalian donor vector with a puromycin resistance marker for transgene integration at the AAVS1 locus in the human genome. Use together with pCas-Guide-AAVS1.

Sequence Author: OriGene

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HomePlasmidsCRISPR PlasmidspAAVS1-Puro-DNR
KflI (7474) AvrII (7363) XcmI (7294) PstI (7133) BbvCI (6704) AleI (6478) FseI (6388) PmeI (6378) FLAG PspXI (6300) NotI (6294) MluI (6285) AscI (6248) MreI - SgrAI (6244) AsiSI (6242) KpnI (6220) Acc65I (6216) AccI (6204) SalI (6203) EcoRI (6197) NcoI (5936) SnaBI (5916) NdeI (5810) CMV enhancer BsmI (5089) PflFI - Tth111I (4936) PasI (4927) BstAPI (4411) BspDI - ClaI (4323) ApaI (3756) Bsu36I (7486) PciI (436) PsiI (2599) PmlI (3305) PshAI (3465) PspOMI (3752) pAAVS1-Puro-DNR 7507 bp
KflI  (7474)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different KflI sites may not be compatible.
AvrII  (7363)
1 site		 C C T A G G G G A T C C
XcmI  (7294)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
PstI  (7133)
1 site		 C T G C A G G A C G T C
BbvCI  (6704)
1 site		 C C T C A G C G G A G T C G
AleI  (6478)
1 site		 C A C N N N N G T G G T G N N N N C A C
FseI  (6388)
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.
PmeI  (6378)
1 site		 G T T T A A A C C A A A T T T G
PspXI  (6300)
1 site		 V C T C G A G B B G A G C T C V
NotI  (6294)
1 site		 G C G G C C G C C G C C G G C G
MluI  (6285)
1 site		 A C G C G T T G C G C A
AscI  (6248)
1 site		 G G C G C G C C C C G C G C G G
MreI  (6244)
1 site		 C G C C G G C G G C G G C C G C
SgrAI  (6244)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
AsiSI  (6242)
1 site		 G C G A T C G C C G C T A G C G
KpnI  (6220)
1 site		 G G T A C C C C A T G G
Acc65I  (6216)
1 site		 G G T A C C C C A T G G
AccI  (6204)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SalI  (6203)
1 site		 G T C G A C C A G C T G
EcoRI  (6197)
1 site		 G A A T T C C T T A A G
NcoI  (5936)
1 site		 C C A T G G G G T A C C
SnaBI  (5916)
1 site		 T A C G T A A T G C A T
NdeI  (5810)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BsmI  (5089)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
PflFI  (4936)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (4936)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PasI  (4927)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
BstAPI  (4411)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
BspDI  (4323)
1 site		 A T C G A T T A G C T A
ClaI  (4323)
1 site		 A T C G A T T A G C T A
ApaI  (3756)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
Bsu36I  (7486)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
PciI  (436)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PsiI  (2599)
1 site		 T T A T A A A A T A T T
PmlI  (3305)
1 site		 C A C G T G G T G C A C
PshAI  (3465)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
PspOMI  (3752)
1 site		 G G G C C C C C C G G G
AmpR
1256 .. 2116  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1256 .. 2047  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1256 .. 2116  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2048 .. 2116  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1256 .. 2116  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
4381 .. 4980  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4381 .. 4980  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
497 .. 1085  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
497 .. 1085  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
HA-R
6972 .. 7501  =  530 bp
right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
HA-R
6972 .. 7501  =  530 bp
right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
HA-L
3278 .. 3779  =  502 bp
left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
HA-L
3278 .. 3779  =  502 bp
left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
PGK promoter
5020 .. 5519  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
5020 .. 5519  =  500 bp
mouse phosphoglycerate kinase 1 promoter
hGH poly(A) signal
6421 .. 6897  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
6421 .. 6897  =  477 bp
human growth hormone polyadenylation signal
f1 ori
2503 .. 2958  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2503 .. 2958  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
5637 .. 5940  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
5637 .. 5940  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
5941 .. 6144  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
5941 .. 6144  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
AmpR promoter
2117 .. 2221  =  105 bp
AmpR promoter
2117 .. 2221  =  105 bp
MCS
6197 .. 6300  =  104 bp
multiple cloning site
MCS
6197 .. 6300  =  104 bp
multiple cloning site
Myc
6303 .. 6332  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
6303 .. 6332  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
6351 .. 6374  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
6351 .. 6374  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
ORF:  4330 .. 5007  =  678 bp
ORF:  225 amino acids  =  23.1 kDa
ORF:  2978 .. 3319  =  342 bp
ORF:  113 amino acids  =  13.1 kDa
ORF:  1386 .. 1652  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1256 .. 2116  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3257 .. 3571  =  315 bp
ORF:  104 amino acids  =  11.4 kDa
ORF:  4292 .. 5239  =  948 bp
ORF:  315 amino acids  =  34.8 kDa
ORF:  4381 .. 4980  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
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Individual Sequences & Maps

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pDCC6
pRGE31
gRNA_AAVS1-T1
pCFD3-dU6-3gRNA
pDD162 (Peft-3_Cas9 + Empty sgRNA)
pRGEB31
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pDR274
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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

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