pFGC-pcoCas9

Binary plant vector for expressing plant codon-optimized Case9 from the 35SPPDK promoter.

Sequence Author: Sheen Lab / Addgene #52256

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AanI (11,623) LB T-DNA repeat EcoRI (9934) PaeR7I - PspXI - XhoI (9928) SnaBI (9924) SwaI (9911) XbaI (9902) AscI (9895) I-PpoI (9882) PacI (9863) I-SceI (9853) I-CeuI (9826) Bsu36I (9822) PI-PspI (9801) SbfI (9751) NruI (9742) AsiSI (9736) SmaI - SrfI (9721) TspMI - XmaI (9719) NOS terminator AanI (6692) NcoI (5003) ATG 2xFLAG SV40 NLS AanI (6670) pFGC-pcoCas9 13,330 bp
AanI  (11,623)
3 sites
T T A T A A A A T A T T
EcoRI  (9934)
1 site
G A A T T C C T T A A G
PaeR7I  (9928)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (9928)
1 site
V C T C G A G B B G A G C T C V
XhoI  (9928)
1 site
C T C G A G G A G C T C
SnaBI  (9924)
1 site
T A C G T A A T G C A T
SwaI  (9911)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
XbaI  (9902)
1 site
T C T A G A A G A T C T
AscI  (9895)
1 site
G G C G C G C C C C G C G C G G
I-PpoI  (9882)
1 site
C T C T C T T A A G G T A G C G A G A G A A T T C C A T C G

I-PpoI is a homing endonuclease that can recognize a variety of similar recognition sequences.
PacI  (9863)
1 site
T T A A T T A A A A T T A A T T
I-SceI  (9853)
1 site
T A G G G A T A A C A G G G T A A T A T C C C T A T T G T C C C A T T A

I-SceI is a homing endonuclease that can recognize a variety of similar recognition sequences.
After cleavage, I-SceI can remain bound to DNA and alter its electrophoretic mobility.
I-CeuI  (9826)
1 site
T A A C T A T A A C G G T C C T A A G G T A G C G A A T T G A T A T T G C C A G G A T T C C A T C G C T

I-CeuI is a homing endonuclease that can recognize a variety of similar recognition sequences.
Bsu36I  (9822)
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.
PI-PspI  (9801)
1 site
T G G C A A A C A G C T A T T A T G G G T A T T A T G G G T A C C G T T T G T C G A T A A T A C C C A T A A T A C C C A

PI-PspI is a homing endonuclease that can recognize a variety of similar recognition sequences.
SbfI  (9751)
1 site
C C T G C A G G G G A C G T C C
NruI  (9742)
1 site
T C G