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Plasmid Files

gRNA_AAVS1-T1

Plasmid for guide RNA (gRNA) targeting of an AAVS1 fragment interrupting an integrated GFP gene in human cells, using the AAVS1 T1 target sequence.

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gRNA_AAVS1-T1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Church Lab / Addgene #41817
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BspLU11I - PciI - PscI (3848) AdeI - DraIII (3045) FseI - RigI (2968) CsiI - MabI - SexAI * (2855) SgrAI (2806) SmaI (2782) Cfr9I - TspMI - XmaI - XmaCI (2780) AatII (2738) ZraI (2736) BsePI - BssHII - MauBI - PauI - PteI (2728) SfiI (2692) TsoI (2349) CpoI - CspI - RsrII - Rsr2I (2336) Bse3DI - BseMI - BsrDI (2053) HindIII (276) Ecl136II - Eco53kI - EcoICRI (290) Psp124BI - SacI - SstI (292) AhlI - BcuI - SpeI (300) FauNDI - NdeI (541) SalI (751) AccI - FblI - XmiI (752) Asp700I - MroXI - PdmI - XmnI (762) PstI (807) Eco32I - EcoRV (810) CciNI - NotI (825) PaeR7I - PspXI - Sfr274I - SlaI - XhoI (831) Bsp120I - PspOMI (849) ApaI (853) BpvUI - Ple19I - PvuI (1004) BsiWI - BstSNI - Eco105I - Pfl23II - PspLI - SnaBI (1034) BsaI - Bso31I - BspTNI - Eco31I (1259) XcmI (1636) BclI * - FbaI * - Ksp22I * (1661) AspI - PflFI - PsyI - Tth111I (1938) gRNA_AAVS1-T1 3973 bp
BspLU11I  (3848)
1 site
A C A T G T T G T A C A

BspLU11I is inhibited by nonionic detergents.
PciI  (3848)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PscI  (3848)
1 site
A C A T G T T G T A C A

PscI is inhibited by nonionic detergents.
AdeI  (3045)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different AdeI sites may not be compatible.
DraIII  (3045)
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.
FseI  (2968)
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.
RigI  (2968)
1 site
G G C C G G C C C C G G C C G G
CsiI  (2855)
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.
MabI  (2855)
1 site
A C C W G G T T G G W C C A

Sticky ends from different MabI sites may not be compatible.
SexAI  (2855)
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.
SgrAI  (2806)
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.
SmaI  (2782)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Cfr9I  (2780)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the Cfr9I recognition sequence.
TspMI  (2780)
1 site
C C C G G G G G G C C C
XmaI  (2780)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
XmaCI  (2780)
1 site
C C C G G G G G G C C C
AatII  (2738)
1 site
G A C G T C C T G C A G
ZraI  (2736)
1 site
G A C G T C C T G C A G
BsePI  (2728)
1 site
G C G C G C C G C G C G
BssHII  (2728)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
MauBI  (2728)
1 site
C G C G C G C G G C G C G C G C
PauI  (2728)
1 site
G C G C G C C G C G C G
PteI  (2728)
1 site
G C G C G C C G C G C G
SfiI  (2692)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
TsoI  (2349)
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).
CpoI  (2336)
1 site
C G G W C C G G C C W G G C

Sticky ends from different CpoI sites may not be compatible.
CspI  (2336)
1 site
C G G W C C G G C C W G G C

Sticky ends from different CspI sites may not be compatible.
RsrII  (2336)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
Rsr2I  (2336)
1 site
C G G W C C G G C C W G G C

Sticky ends from different Rsr2I sites may not be compatible.
Bse3DI  (2053)
1 site
G C A A T G N N C G T T A C

Sticky ends from different Bse3DI sites may not be compatible.
BseMI  (2053)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BseMI sites may not be compatible.
BsrDI  (2053)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
HindIII  (276)
1 site
A A G C T T T T C G A A
Ecl136II  (290)
1 site
G A G C T C C T C G A G
Eco53kI  (290)
1 site
G A G C T C C T C G A G
EcoICRI  (290)
1 site
G A G C T C C T C G A G
Psp124BI  (292)
1 site
G A G C T C C T C G A G
SacI  (292)
1 site
G A G C T C C T C G A G
SstI  (292)
1 site
G A G C T C C T C G A G
AhlI  (300)
1 site
A C T A G T T G A T C A
BcuI  (300)
1 site
A C T A G T T G A T C A
SpeI  (300)
1 site
A C T A G T T G A T C A
FauNDI  (541)
1 site
C A T A T G G T A T A C
NdeI  (541)
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.
SalI  (751)
1 site
G T C G A C C A G C T G
AccI  (752)
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.
FblI  (752)
1 site
G T M K A C C A K M T G

Sticky ends from different FblI sites may not be compatible.
XmiI  (752)
1 site
G T M K A C C A K M T G

Sticky ends from different XmiI sites may not be compatible.
Asp700I  (762)
1 site
G A A N N N N T T C C T T N N N N A A G
MroXI  (762)
1 site
G A A N N N N T T C C T T N N N N A A G
PdmI  (762)
1 site
G A A N N N N T T C C T T N N N N A A G
XmnI  (762)
1 site
G A A N N N N T T C C T T N N N N A A G
PstI  (807)
1 site
C T G C A G G A C G T C
Eco32I  (810)
1 site
G A T A T C C T A T A G
EcoRV  (810)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
CciNI  (825)
1 site
G C G G C C G C C G C C G G C G
NotI  (825)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (831)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (831)
1 site
V C T C G A G B B