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Plasmid Files

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.

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pAAVS1-Puro-DNR.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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KflI (7474) AvrII (7363) XcmI (7294) PstI (7133) BbvCI (6704) AleI (6478) FseI (6388) PmeI (6378) 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) 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

PmlI gradually loses activity when stored at -20°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
   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
   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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