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pROSA26-Puro-DNR

Mammalian donor vector with a puromycin resistance marker for transgene integration at the ROSA26 locus in the mouse genome. Use together with pCas‑Guide-ROSA26.

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pROSA26-Puro-DNR.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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BsrGI (7805) MfeI (7149) BbvCI (6872) AleI (6646) FseI (6556) PmeI (6546) PspXI (6468) NotI (6462) MluI (6453) BmtI (6445) NheI (6441) AscI (6416) MreI - SgrAI (6412) AsiSI (6410) KpnI (6388) Acc65I (6384) BamHI (6378) AccI (6372) SalI (6371) EcoRI (6365) SacI (6312) Eco53kI (6310) NcoI (6104) SnaBI (6084) NdeI (5978) PpuMI (5269) BfuAI - BspMI (5173) PflFI - Tth111I (5104) BstEII (5008) BstAPI (4579) BspDI - ClaI (4491) PflMI (4146) ApaI (3924) BspQI - SapI (319) PciI (435) SbfI (3787) PspOMI (3920) pROSA26-Puro-DNR 7810 bp
BsrGI  (7805)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
MfeI  (7149)
1 site
C A A T T G G T T A A C
BbvCI  (6872)
1 site
C C T C A G C G G A G T C G
AleI  (6646)
1 site
C A C N N N N G T G G T G N N N N C A C
FseI  (6556)
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  (6546)
1 site
G T T T A A A C C A A A T T T G
PspXI  (6468)
1 site
V C T C G A G B B G A G C T C V
NotI  (6462)
1 site
G C G G C C G C C G C C G G C G
MluI  (6453)
1 site
A C G C G T T G C G C A
BmtI  (6445)
1 site
G C T A G C C G A T C G
NheI  (6441)
1 site
G C T A G C C G A T C G
AscI  (6416)
1 site
G G C G C G C C C C G C G C G G
MreI  (6412)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (6412)
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  (6410)
1 site
G C G A T C G C C G C T A G C G
KpnI  (6388)
1 site
G G T A C C C C A T G G
Acc65I  (6384)
1 site
G G T A C C C C A T G G
BamHI  (6378)
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.
AccI  (6372)
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  (6371)
1 site
G T C G A C C A G C T G
EcoRI  (6365)
1 site
G A A T T C C T T A A G
SacI  (6312)
1 site
G A G C T C C T C G A G
Eco53kI  (6310)
1 site
G A G C T C C T C G A G
NcoI  (6104)
1 site
C C A T G G G G T A C C
SnaBI  (6084)
1 site
T A C G T A A T G C A T
NdeI  (5978)
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.
PpuMI  (5269)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BfuAI  (5173)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (5173)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PflFI  (5104)
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  (5104)
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.
BstEII  (5008)
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.
BstAPI  (4579)
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  (4491)
1 site
A T C G A T T A G C T A
ClaI  (4491)
1 site
A T C G A T T A G C T A
PflMI  (4146)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
ApaI  (3924)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BspQI  (319)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (319)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PciI  (435)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SbfI  (3787)
1 site
C C T G C A G G G G A C G T C C
PspOMI  (3920)
1 site
G G G C C C C C C G G G
AmpR
1255 .. 2115  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1255 .. 2046  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1255 .. 2115  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2047 .. 2115  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1255 .. 2115  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
HA-R
7161 .. 7810  =  650 bp
right homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)
HA-R
7161 .. 7810  =  650 bp
right homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)
HA-L
3271 .. 3919  =  649 bp
left homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)
HA-L
3271 .. 3919  =  649 bp
left homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)
PuroR
4549 .. 5148  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4549 .. 5148  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
496 .. 1084  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
496 .. 1084  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
5188 .. 5687  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
5188 .. 5687  =  500 bp
mouse phosphoglycerate kinase 1 promoter
hGH poly(A) signal
6589 .. 7065  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
6589 .. 7065  =  477 bp
human growth hormone polyadenylation signal
f1 ori
2502 .. 2957  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2502 .. 2957  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
5805 .. 6108  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
5805 .. 6108  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
6109 .. 6312  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
6109 .. 6312  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
MCS
6364 .. 6558  =  195 bp
multiple cloning site
MCS
6364 .. 6558  =  195 bp
multiple cloning site
AmpR promoter
2116 .. 2220  =  105 bp
AmpR promoter
2116 .. 2220  =  105 bp
T7 promoter
6338 .. 6356  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
6338 .. 6356  =  19 bp
promoter for bacteriophage T7 RNA polymerase
Myc
6471 .. 6500  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
Myc
6471 .. 6500  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
FLAG
6519 .. 6542  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
6519 .. 6542  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
ORF:  2977 .. 3405  =  429 bp
ORF:  142 amino acids  =  15.8 kDa
ORF:  4498 .. 5175  =  678 bp
ORF:  225 amino acids  =  23.1 kDa
ORF:  1385 .. 1651  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  4460 .. 5407  =  948 bp
ORF:  315 amino acids  =  34.8 kDa
ORF:  1255 .. 2115  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4549 .. 5148  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
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