Resources
Plasmid Files

pGuide-it-sgRNA1 Vector (Linear)

Linearized CRISPR/Cas9 vector with an improved scaffold, for cloning and expression of a single guide RNA (sgRNA).

To obtain this DNA and protein sequence with restriction sites, please download
 SnapGene or the free  SnapGene Viewer.

pGuide-it-sgRNA1 Vector (Linear).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
Download Free Trial Get SnapGene Viewer


End (3020) Guide-it™ Sequencing Primer 1 (2943 .. 2966) BsaAI - BstBAI - Ppu21I (2885) BglII (2761) AxyI - Bse21I - Bsu36I - Eco81I (2743) AcsI - ApoI - EcoRI - XapI (2732) BbeI - PluTI (2575) DinI - EgeI - EheI - SfoI (2573) Mly113I - NarI (2572) KasI - SspDI (2571) BstAPI (2521) PfoI (2382) AatII (2271) ZraI (2269) SspI (2153) Asp700I - MroXI - PdmI - XmnI (1948) Acc113I - BmcAI - ScaI - ZrmI (1829) TsoI (1748) NmeAIII (1497) Start (0) HindIII (97) BspQI - LguI - PciSI - SapI (340) AflIII - BspLU11I - PciI - PscI (456) BseYI (760) GsaI - PspFI (764) AlwNI - CaiI (872) AhdI - AspEI - BmeRI - DriI - Eam1105I (1349) BsaI - Bso31I - BspTNI - Eco31I (1410) BpmI - GsuI (1419) Bse118I - BsrFI - BssAI - Cfr10I (1429) pGuide-it-sgRNA1 Vector (Linear) 3020 bp
End  (3020)
0 sites
BsaAI  (2885)
1 site
Y A C G T R R T G C A Y
BstBAI  (2885)
1 site
Y A C G T R R T G C A Y
Ppu21I  (2885)
1 site
Y A C G T R R T G C A Y
BglII  (2761)
1 site
A G A T C T T C T A G A
AxyI  (2743)
1 site
C C T N A G G G G A N T C C

Sticky ends from different AxyI sites may not be compatible.
Bse21I  (2743)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bse21I sites may not be compatible.
Bsu36I  (2743)
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.
Eco81I  (2743)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Eco81I sites may not be compatible.
AcsI  (2732)
1 site
R A A T T Y Y T T A A R
ApoI  (2732)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (2732)
1 site
G A A T T C C T T A A G
XapI  (2732)
1 site
R A A T T Y Y T T A A R
BbeI  (2575)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the BbeI recognition sequence.
PluTI  (2575)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
DinI  (2573)
1 site
G G C G C C C C G C G G
EgeI  (2573)
1 site
G G C G C C C C G C G G
EheI  (2573)
1 site
G G C G C C C C G C G G
SfoI  (2573)
1 site
G G C G C C C C G C G G
Mly113I  (2572)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the Mly113I recognition sequence.
NarI  (2572)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (2571)
1 site
G G C G C C C C G C G G
SspDI  (2571)
1 site
G G C G C C C C G C G G
BstAPI  (2521)
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.
PfoI  (2382)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
AatII  (2271)
1 site
G A C G T C C T G C A G
ZraI  (2269)
1 site
G A C G T C C T G C A G
SspI  (2153)
1 site
A A T A T T T T A T A A
Asp700I  (1948)
1 site
G A A N N N N T T C C T T N N N N A A G
MroXI  (1948)
1 site
G A A N N N N T T C C T T N N N N A A G
PdmI  (1948)
1 site
G A A N N N N T T C C T T N N N N A A G
XmnI  (1948)
1 site
G A A N N N N T T C C T T N N N N A A G
Acc113I  (1829)
1 site
A G T A C T T C A T G A
BmcAI  (1829)
1 site
A G T A C T T C A T G A
ScaI  (1829)
1 site
A G T A C T T C A T G A
ZrmI  (1829)
1 site
A G T A C T T C A T G A
TsoI  (1748)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
NmeAIII  (1497)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
Start  (0)
0 sites
HindIII  (97)
1 site
A A G C T T T T C G A A
BspQI  (340)
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.
LguI  (340)
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 LguI sites may not be compatible.
PciSI  (340)
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 PciSI sites may not be compatible.
SapI  (340)
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.
AflIII  (456)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
BspLU11I  (456)
1 site
A C A T G T T G T A C A

BspLU11I is inhibited by nonionic detergents.
PciI  (456)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PscI  (456)
1 site
A C A T G T T G T A C A

PscI is inhibited by nonionic detergents.
BseYI  (760)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
GsaI  (764)
1 site
C C C A G C G G G T C G
PspFI  (764)
1 site
C C C A G C G G G T C G
AlwNI  (872)
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.
CaiI  (872)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different CaiI sites may not be compatible.
AhdI  (1349)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AspEI  (1349)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AspEI may be hard to ligate.
Sticky ends from different AspEI sites may not be compatible.
BmeRI  (1349)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by BmeRI may be hard to ligate.
Sticky ends from different BmeRI sites may not be compatible.
DriI  (1349)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by DriI may be hard to ligate.
Sticky ends from different DriI sites may not be compatible.
Eam1105I  (1349)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by Eam1105I may be hard to ligate.
Sticky ends from different Eam1105I sites may not be compatible.
BsaI  (1410)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
Bso31I  (1410)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different Bso31I sites may not be compatible.
BspTNI  (1410)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BspTNI sites may not be compatible.
Eco31I  (1410)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different Eco31I sites may not be compatible.
BpmI  (1419)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
GsuI  (1419)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Sticky ends from different GsuI sites may not be compatible.
After cleavage, GsuI can remain bound to DNA and alter its electrophoretic mobility.
GsuI is typically used at 30°C, but is 70% active at 37°C.
For full activity, add fresh S-adenosylmethionine (SAM).
Bse118I  (1429)
1 site
R C C G G Y Y G G C C R
BsrFI  (1429)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
BssAI  (1429)
1 site
R C C G G Y Y G G C C R
Cfr10I  (1429)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the Cfr10I recognition sequence.
Guide-it™ Sequencing Primer 1
24-mer  /  38% GC
1 binding site
2943 .. 2966  =  24 annealed bases
Tm  =  56°C
AmpR
1276 .. 2136  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1276 .. 2067  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1276 .. 2136  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2068 .. 2136  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1276 .. 2136  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
517 .. 1105  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
517 .. 1105  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
U6 promoter
2776 .. 3016  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2776 .. 3016  =  241 bp
RNA polymerase III promoter for human U6 snRNA
AmpR promoter
2137 .. 2241  =  105 bp
AmpR promoter
2137 .. 2241  =  105 bp
gRNA scaffold
1 .. 86  =  86 bp
improved guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
1 .. 86  =  86 bp
improved guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
ORF:  1406 .. 1672  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2522 .. 2746  =  225 bp
ORF:  74 amino acids  =  8.8 kDa
ORF:  1276 .. 2136  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps