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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HomePlasmidsCRISPR PlasmidspFGC-pcoCas9
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 nucleoplasmin NLS 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 C G A A G C G C T
AsiSI  (9736)
1 site		 G C G A T C G C C G C T A G C G
SmaI  (9721)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
SrfI  (9721)
1 site		 G C C C G G G C C G G G C C C G
TspMI  (9719)
1 site		 C C C G G G G G G C C C
XmaI  (9719)
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.
AanI  (6692)
3 sites		 T T A T A A A A T A T T
NcoI  (5003)
1 site		 C C A T G G G G T A C C
AanI  (6670)
3 sites		 T T A T A A A A T A T T
ATG
5005 .. 5007  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5005 .. 5007  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
2xFLAG
5008 .. 5055  =  48 bp
16 amino acids  =  2.0 kDa
Product: two tandem FLAG® epitope tags
2xFLAG
5008 .. 5055  =  48 bp
16 amino acids  =  2.0 kDa
Product: two tandem FLAG® epitope tags
SV40 NLS
5062 .. 5082  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5062 .. 5082  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
pcoCas9
5107 .. 9396  =  4290 bp
1367 amino acids  =  158.3 kDa
2 segments  =  4101 bp
Product: Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
plant codon-optimized Cas9 gene containing the potato IV2 intron
pcoCas9
5107 .. 9396  =  4290 bp
1367 amino acids  =  158.3 kDa
2 segments  =  4101 bp
   Segment 1:  
   5107 .. 6543  =  1437 bp
   479 amino acids  =  55.4 kDa
Product: Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
plant codon-optimized Cas9 gene contain