pEarleyGate 303

Gateway®-compatible promoterless plant transformation vector with a C-terminal Myc tag.

Sequence Author: Pikaard Lab

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AseI (94) BclI * (10,145) PsiI (10,014) BspHI (9688) EcoRV (9265) NsiI (9154) PluTI (8006) SfoI (8004) NarI (8003) KasI (8002) EcoNI (7530) BseRI (7494) BmtI (7221) NheI (7217) BsiWI (7101) XmnI (95) Eco53kI (103) SacI (105) FspAI (523) PaqCI (587) AfeI (760) attR1 BspEI (1806) NcoI (2111) MluI (2257) BbvCI (2535) SrfI (2681) BstXI (2798) Myc BglII (3093) XbaI (3112) PacI (3125) SpeI (3137) MfeI (3441) BstEII (3713) AhdI (3843) SbfI (3868) HindIII (3876) PvuI (4002) PmeI (4089) pEarleyGate 303 10,382 bp
AseI  (94)
1 site
A T T A A T T A A T T A
BclI  (10,145)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
PsiI  (10,014)
1 site
T T A T A A A A T A T T
BspHI  (9688)
1 site
T C A T G A A G T A C T
EcoRV  (9265)
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.
NsiI  (9154)
1 site
A T G C A T T A C G T A
PluTI  (8006)
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.
SfoI  (8004)
1 site
G G C G C C C C G C G G
NarI  (8003)
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  (8002)
1 site
G G C G C C C C G C G G
EcoNI  (7530)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BseRI  (7494)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BmtI  (7221)
1 site
G C T A G C C G A T C G
NheI  (7217)
1 site
G C T A G C C G A T C G
BsiWI  (7101)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
XmnI  (95)
1 site
G A A N N N N T T C C T T N N N N A A G
Eco53kI  (103)
1 site
G A G C T C C T C G A G
SacI  (105)
1 site
G A G C T C C T C G A G
FspAI  (523)
1 site
R T G C G C A Y Y A C G C G T R
PaqCI  (587)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
AfeI  (760)
1 site
A G C G C T T C G C G A
BspEI  (1806)
1 site
T C C G G A A G G C C T
NcoI  (2111)
1 site
C C A T G G G G T A C C
MluI  (2257)
1 site
A C G C G T T G C G C A