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pEarleyGate 202

Gateway®-compatible plant transformation vector with an N-terminal FLAG tag.

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pEarleyGate 202.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Pikaard Lab
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BclI * (11,539) PsiI (11,408) BspHI (11,082) EcoRV (10,659) PluTI (9400) SfoI (9398) NarI (9397) KasI (9396) EcoNI (8924) BmtI (8615) NheI (8611) BsiWI (8495) PmeI (5483) AseI (92) Acc65I (407) KpnI (411) FspAI (521) AarI (585) AfeI (758) BstBI (1770) StuI (1868) PshAI (2467) PaeR7I - PspXI - XhoI (2654) BspEI (3209) BbvCI (3938) SrfI (4084) BstXI (4201) AvrII (4473) PacI (4519) SpeI (4531) MfeI (4835) BstEII (5107) AhdI (5237) SbfI (5262) HindIII (5270) PvuI (5396) pEarleyGate 202 11,776 bp
BclI  (11,539)
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  (11,408)
1 site
T T A T A A A A T A T T
BspHI  (11,082)
1 site
T C A T G A A G T A C T
EcoRV  (10,659)
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.
PluTI  (9400)
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  (9398)
1 site
G G C G C C C C G C G G
NarI  (9397)
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  (9396)
1 site
G G C G C C C C G C G G
EcoNI  (8924)
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.
BmtI  (8615)
1 site
G C T A G C C G A T C G
NheI  (8611)
1 site
G C T A G C C G A T C G
BsiWI  (8495)
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.
PmeI  (5483)
1 site
G T T T A A A C C A A A T T T G
AseI  (92)
1 site
A T T A A T T A A T T A
Acc65I  (407)
1 site
G G T A C C C C A T G G
KpnI  (411)
1 site
G G T A C C C C A T G G
FspAI  (521)
1 site
R T G C G C A Y Y A C G C G T R
AarI  (585)
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 AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
AfeI  (758)
1 site