pHELLSGATE 12

High-throughput binary Gateway® vector with two introns, for silencing genes in plants using intron-containing hairpin RNA.
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AgeI (17,247) XbaI (16,903) XbaI (15,484) BspDI - ClaI (15,262) PacI (15,151) PDK intron KpnI (14,465) Acc65I (14,461) MCS 1 EcoRI (14,455) XhoI (14,449) XhoI (13,020) ScaI (12,386) StuI (12,234) SacI (11,646) Eco53kI (11,644) PmeI (11,220) BsiWI (11,055) FseI (10,595) Bsu36I (330) RsrII (1119) AflII (1923) MauBI (2118) LB T-DNA repeat BspEI (3460) AleI (5445) SexAI * (5533) FspAI (6101) NruI (6598) pHELLSGATE 12 17,681 bp
AgeI  (17,247)
1 site
A C C G G T T G G C C A
XbaI  (16,903)
2 sites
T C T A G A A G A T C T
XbaI  (15,484)
2 sites
T C T A G A A G A T C T
BspDI  (15,262)
1 site
A T C G A T T A G C T A
ClaI  (15,262)
1 site
A T C G A T T A G C T A
PacI  (15,151)
1 site
T T A A T T A A A A T T A A T T
KpnI  (14,465)
1 site
G G T A C C C C A T G G
Acc65I  (14,461)
1 site
G G T A C C C C A T G G
EcoRI  (14,455)
1 site
G A A T T C C T T A A G
XhoI  (14,449)
2 sites
C T C G A G G A G C T C
XhoI  (13,020)
2 sites
C T C G A G G A G C T C
ScaI  (12,386)
1 site
A G T A C T T C A T G A
StuI  (12,234)
1 site
A G G C C T T C C G G A
SacI  (11,646)
1 site
G A G C T C C T C G A G
Eco53kI  (11,644)
1 site
G A G C T C C T C G A G
PmeI  (11,220)
1 site
G T T T A A A C C A A A T T T G
BsiWI  (11,055)
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.
FseI  (10,595)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
Bsu36I  (330)
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.
RsrII  (1119)
1 site
C G G W C