pEarleyGate 100

Gateway®-compatible plant transformation vector with no encoded tag.

Sequence Author: Pikaard Lab

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LB T-DNA repeat BclI * (11,412) PsiI (11,281) BspHI (10,955) EcoRV (10,532) PluTI (9273) SfoI (9271) NarI (9270) KasI (9269) EcoNI (8797) BmtI (8488) NheI (8484) BsiWI (8368) AseI (92) Eco53kI (101) SacI (103) Acc65I (407) KpnI (411) FspAI (521) PaqCI (585) AfeI (758) BstBI (1770) StuI (1868) PshAI (2467) PaeR7I - XhoI (2654) BspEI (3105) NcoI (3410) BbvCI (3834) SrfI (3980) BstXI (4097) AvrII (4369) XbaI (4379) PacI (4392) SpeI (4404) MfeI (4708) BstEII (4980) AhdI (5110) SbfI (5135) HindIII (5143) PvuI (5269) PmeI (5356) pEarleyGate 100 11,649 bp
BclI  (11,412)
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  (11,281)
1 site
T T A T A A A A T A T T
BspHI  (10,955)
1 site
T C A T G A A G T A C T
EcoRV  (10,532)
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  (9273)
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  (9271)
1 site
G G C G C C C C G C G G
NarI  (9270)
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  (9269)
1 site
G G C G C C C C G C G G
EcoNI  (8797)
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  (8488)
1 site
G C T A G C C G A T C G
NheI  (8484)
1 site
G C T A G C C G A T C G
BsiWI  (8368)
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.
AseI  (92)
1 site