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Plasmid Files

pNW3

Plasmid for mammalian cell expression of the Csy4 CRISPR-associated endoribonuclease and a D10A nickase mutant of Cas9.

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pNW3.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Joung Lab / Addgene #53372
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SexAI * (9650) ScaI (9216) PvuI (9106) BsaI (8797) AhdI (8736) AvrII (7510) SfiI (7463) HindIII (7139) BstXI (6652) NotI (6592) PaeR7I - XhoI (6514) BmtI (6030) NheI (6026) FspAI (5334) SpeI (18) BlpI (1154) MreI - SgrAI (1311) XbaI (1623) Acc65I (1735) KpnI (1739) TspMI - XmaI (1746) SmaI (1748) AgeI (1756) BsmBI (2240) HpaI (2349) BstZ17I (2598) BmgBI (2603) BstEII (3098) BsiWI (3192) BsrGI (3942) EcoRV (4225) PshAI (4366) PvuII (4468) pNW3 9654 bp
SexAI  (9650)
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.
ScaI  (9216)
1 site
A G T A C T T C A T G A
PvuI  (9106)
1 site
C G A T C G G C T A G C
BsaI  (8797)
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.
AhdI  (8736)
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.
AvrII  (7510)
1 site
C C T A G G G G A T C C
SfiI  (7463)
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.
HindIII  (7139)
1 site
A A G C T T T T C G A