pGreenII 0229

Agrobacterium binary vector with kanamycin-and bialophos/phosphinothricin-resistance genes.

Sequence Author: pGreen website

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DraIII (4322) BsrDI (4298) NruI (4131) TsoI (3949) PasI (3913) EcoNI (3875) AsiSI (3790) Bpu10I (3768) PflMI (3528) AlwNI (3001) ApaLI (2899) PspFI (2893) BseYI (2889) BglII (2580) StuI (2527) BspQI - SapI (2403) lac operator SacI (2124) StyI (421) FseI (528) AanI - PsiI (546) BglII (627) LB T-DNA repeat MauBI (702) AflIII - PciI (816) AflII (896) BmgBI (1021) FspAI (1098) AarI - BfuAI - BspMI (1162) PpuMI - SanDI (1275) AfeI (1335) PflFI - Tth111I (1352) BsgI (1455) BsaHI (1484) NheI (1605) BmtI (1609) Bsu36I (1621) BlpI (1666) Acc65I (2018) KpnI (2022) PspOMI (2024) ApaI (2028) AbsI - PaeR7I - PspXI - XhoI (2033) SalI (2039) AccI (2040) HindIII (2054) EcoRV (2062) EcoRI (2066) PstI (2076) TspMI - XmaI (2078) SmaI (2080) BamHI (2084) SpeI (2090) XbaI (2096) NotI (2103) AleI - MslI (2114) SacII (2115) BstXI (2116) Eco53kI (2122) pGreenII 0229 4448 bp
DraIII  (4322)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BsrDI  (4298)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
NruI  (4131)
1 site
T C G C G A A G C G C T
TsoI  (3949)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PasI  (3913)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
EcoNI  (3875)
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.
AsiSI  (3790)
1 site
G C G A T C G C C G C T A G C G
Bpu10I  (3768)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
PflMI  (3528)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
AlwNI  (3001)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
ApaLI  (2899)
1 site
G T G C A C C A C G T G
PspFI  (2893)
1 site
C C C A G C G G G T C G
BseYI  (2889)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
BglII  (2580)
2 sites
A G A T C T T C T A G A
StuI  (2527)
1 site
A G G C C T T C C G G A
BspQI  (2403)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2403)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
SacI  (2124)
1 site
G A G C T C C T C G A G
StyI  (421)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
FseI  (528)
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.
AanI  (546)
1 site
T T A T A A A A T A T T
PsiI  (546)
1 site
T T A T A A A A T A T T
BglII  (627)
2 sites
A G A T C T T C T A G A
MauBI  (702)
1 site
C G C G C G C G G C G C G C G C
AflIII  (816)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (816)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AflII  (896)
1 site
C T T A A G G A A T T C
BmgBI  (1021)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
FspAI  (1098)
1 site
R T G C G C A Y Y A C G C G T R
AarI  (1162)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BfuAI  (1162)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1162)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PpuMI  (1275)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
SanDI  (1275)
1 site
G G G W C C C C C C W G G G

Sticky ends from different SanDI sites may not be compatible.
AfeI  (1335)
1 site
A G C G C T T C G C G A
PflFI  (1352)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1352)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsgI  (1455)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaHI  (1484)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
NheI  (1605)
1 site
G C T A G C C G A T C G
BmtI  (1609)
1 site
G C T A G C C G A T C G
Bsu36I  (1621)
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.
BlpI  (1666)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
Acc65I  (2018)
1 site
G G T A C C C C A T G G
KpnI  (2022)
1 site
G G T A C C C C A T G G
PspOMI  (2024)
1 site
G G G C C C C C C G G G
ApaI  (2028)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AbsI  (2033)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2033)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2033)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2033)
1 site
C T C G A G G A G C T C
SalI  (2039)
1 site
G T C G A C C A G C T G
AccI  (2040)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HindIII  (2054)
1 site
A A G C T T T T C G A A
EcoRV  (2062)
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.
EcoRI  (2066)
1 site
G A A T T C C T T A A G
PstI  (2076)
1 site
C T G C A G G A C G T C
TspMI  (2078)
1 site
C C C G G G G G G C C C
XmaI  (2078)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2080)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (2084)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
SpeI  (2090)
1 site
A C T A G T T G A T C A