pEarleyGate 205

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

Sequence Author: Pikaard Lab

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BclI * (11,951) PsiI (11,820) BspHI (11,494) EcoRV (11,071) EcoNI (9336) BsiWI (8907) PmeI (5895) LB T-DNA repeat Acc65I (407) KpnI (411) FspAI (521) PaqCI (585) BstBI (1770) StuI (1868) PshAI (2467) PaeR7I - XhoI (2654) attR1 lac UV5 promoter BspEI (3105) NcoI (3410) BbvCI (3834) SrfI (3980) BstXI (4097) attR2 AvrII (4369) TEV Site ProtA ProtA PacI (4931) SpeI (4943) MfeI (5247) BstEII (5519) AhdI (5649) SbfI (5674) HindIII (5682) PvuI (5808) pEarleyGate 205 12,188 bp
BclI  (11,951)
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  (11,820)
1 site
T T A T A A A A T A T T
BspHI  (11,494)
1 site
T C A T G A A G T A C T
EcoRV  (11,071)
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.
EcoNI  (9336)
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.
BsiWI  (8907)
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  (5895)
1 site
G T T T A A A C C A A A T T T G
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
PaqCI  (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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
BstBI  (1770)
1 site
T T C G A A A A G C T T
StuI  (1868)
1 site
A G G C C T T C C G G A
PshAI  (2467)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
PaeR7I  (2654)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2654)
1 site
C T C G A G G A G C T C
BspEI  (3105)
1 site
T C C G G A A G G C C T
NcoI  (3410)
1 site
C C A T G G G G T A C C
BbvCI  (3834)
1 site
C C T C A G C G G A G T C G
SrfI  (3980)
1 site
G C C C G G G C C G G G C C C G
BstXI  (4097)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
AvrII  (4369)
1 site
C C T A G G G G A T C C
PacI  (4931)
1 site
T T A A T T A A A A T T A A T T
SpeI  (4943)
1 site
A C T A G T T G A T C A
MfeI  (5247)
1 site
C A A T T G G T T A A C
BstEII  (5519)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
AhdI  (5649)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
SbfI  (5674)
1 site
C C T G C A G G G G A C G T C C
HindIII  (5682)
1 site
A A G C T T T T C G A A
PvuI  (5808)
1 site
C G A T C G G C T A G C
pVS1 RepA
8292 .. 9365  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
pVS1 RepA
8292 .. 9365  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
KanR
10,970 .. 11,764  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
10,970 .. 11,764  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
OCS terminator
4959 .. 5666  =  708 bp
octopine synthase terminator
OCS terminator
4959 .. 5666  =  708 bp
octopine synthase terminator
CmR
2896 .. 3555  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2896 .. 3555  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
pVS1 StaA
7234 .. 7863  =  630 bp
209 amino acids  =  22.1 kDa