pCas-Guide-ROSA26

Mammalian guide RNA (gRNA) vector for transgene integration at the ROSA26 locus in the mouse genome. Use together with pROSA26-Puro-DNR.

Sequence Author: OriGene

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Explore Over 2.7k Plasmids: CRISPR Plasmids | More Plasmid Sets
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T7 promoter ScaI (7512) AlwNI (6562) HaeII (6394) BciVI (6349) PciI (6174) BstXI (5898) NsiI (5782) BbvCI (5747) SmaI (5463) TspMI - XmaI (5461) SacII (5435) FseI (5431) NaeI (5429) NgoMIV (5427) PmeI (5421) FLAG PaeR7I - PspXI - XhoI (5343) BsiWI (5331) SV40 NLS SV40 NLS BstEII (5271) XcmI (4712) KflI (4550) BsmI (4145) HpaI (7995) DraIII - PmlI (40) ROSA26 target sequence polIII terminator BsrGI (445) SpeI (501) BtgZI (836) SnaBI (842) Eco53kI (1068) SacI (1070) T7 promoter SalI (1129) Acc65I (1142) KpnI (1146) BglII (1152) AsiSI (1168) BstZ17I (1387) BfuAI - BspMI (2241) PshAI (2748) CsiI - SexAI * (3744) PflFI - Tth111I (3746) pCas-Guide-ROSA26 7998 bp
ScaI  (7512)
1 site
A G T A C T T C A T G A
AlwNI  (6562)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
HaeII  (6394)
1 site
R G C G C Y Y C G C G R
BciVI  (6349)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
PciI  (6174)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstXI  (5898)
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.
NsiI  (5782)
1 site
A T G C A T T A C G T A
BbvCI  (5747)
1 site
C C T C A G C G G A G T C G
SmaI  (5463)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (5461)
1 site
C C C G G G G G G C C C
XmaI  (5461)
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.
SacII  (5435)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
FseI  (5431)
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.
NaeI  (5429)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (5427)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
PmeI  (5421)
1 site
G T T T A A A C C A A A T T T G
PaeR7I  (5343)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5343)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5343)
1 site
C T C G A G G A G C T C
BsiWI  (5331)
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.
BstEII  (5271)
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.
XcmI  (4712)
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.
KflI  (4550)
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.
BsmI  (4145)
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.
HpaI  (7995)
1 site
G T T A A C C A A T T G
DraIII  (40)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
PmlI  (40)
1 site
C A C G T G G T G C A C
BsrGI  (445)
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.
SpeI  (501)
1 site
A C T A G T T G A T C A
BtgZI  (836)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
SnaBI  (842)
1 site
T A C G T A A T G C A T
Eco53kI  (1068)
1 site
G A G C T C C T C G A G
SacI  (1070)
1 site
G A G C T C C T C G A G
SalI  (1129)
1 site
G T C G A C C A G C T G
Acc65I  (1142)
1 site
G G T A C C C C A T G G
KpnI  (1146)
1 site
G G T A C C C C A T G G
BglII  (1152)
1 site
A G A T C T T C T A G A
AsiSI  (1168)
1 site
G C G A T C G C C G C T A G C G
BstZ17I  (1387)
1 site
G T A T A C C A T A T G
BfuAI  (2241)
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  (2241)
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.
PshAI  (2748)
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.
CsiI  (3744)
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  (3744)
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.
PflFI  (3746)
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  (3746)
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.
Cas9
1173 .. 5276  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
1173 .. 5276  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SV40 NLS
5277 .. 5297  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5277 .. 5297  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5304 .. 5324  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5304 .. 5324  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Myc
5346 .. 5375  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
Myc
5346 .. 5375  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
FLAG
5394 .. 5417  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
5394 .. 5417  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
AmpR
6959 .. 7819  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6959 .. 7750  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6959 .. 7819  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7751 .. 7819  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6959 .. 7819  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
hGH poly(A) signal
5464 .. 6086  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
5464 .. 6086  =  623 bp
human growth hormone polyadenylation signal
ori
6207 .. 6795  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6207 .. 6795  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
487 .. 866  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
487 .. 866  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
90 .. 330  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
90 .. 330  =  241 bp
RNA polymerase III promoter for human U6 snRNA
CMV promoter
867 .. 1070  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
867 .. 1070  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
gRNA scaffold
360 .. 435  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
360 .. 435  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
ROSA26 target sequence
340 .. 359  =  20 bp
ROSA26 target sequence
340 .. 359  =  20 bp
T7 promoter
1096 .. 1114  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1096 .. 1114  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
7955 .. 7973  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
7955 .. 7973  =  19 bp
promoter for bacteriophage T7 RNA polymerase
polIII terminator
436 .. 441  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
436 .. 441  =  6 bp
RNA polymerase III transcription terminator
ORF:  6176 .. 6532  =  357 bp
ORF:  118 amino acids  =  13.2 kDa
ORF:  1026 .. 5423  =  4398 bp
ORF:  1465 amino acids  =  169.8 kDa
ORF:  7089 .. 7355  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1431 .. 1673  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
ORF:  3225 .. 3569  =  345 bp
ORF:  114 amino acids  =  13.5 kDa
ORF:  3723 .. 4127  =  405 bp
ORF:  134 amino acids  =  15.6 kDa
ORF:  4596 .. 5219  =  624 bp
ORF:  207 amino acids  =  24.3 kDa
ORF:  5828 .. 6052  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  6959 .. 7819  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pCas-Guide-ROSA26.dna file

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