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Plasmid Files

pGreenII

Improved compact Agrobacterium binary vector with a kanamycin-resistance gene. Also known as pGreenII 0000.

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pGreenII Sequence and MappGreenII.dna
Map and Sequence File   
Sequence Author:  pGreen website
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 BsaBI * (3275) DraIII (3087) BsrDI (3063) BspHI (3031) NruI (2896) TsoI (2714) PasI (2678) EcoNI (2640) SspI (2628) AsiSI (2555) Bpu10I - BsmBI (2533) PflMI (2293) BspHI (2070) AcuI (1898) AlwNI (1766) ApaLI (1664) PspFI (1658) StyI (330) NgoMIV (433) NaeI (435) FseI (437) AanI - AanI - PsiI (455) BglII (536) HpaI (568) FspI (612) BmrI (713) Acc65I (783) KpnI (787) PspOMI (789) EcoO109I (790) ApaI (793) AbsI - PaeR7I - PspXI - XhoI (798) SalI (804) AccI (805) BspDI - ClaI (814) HindIII (819) EcoRV (827) EcoRI (831) PstI (841) TspMI - XmaI (843) SmaI (845) BamHI (849) SpeI (855) XbaI (861) NotI (868) BpmI (873) AleI - MslI (879) SacII (880) BstXI (881) Eco53kI (887) SacI (889) lac operator BspQI - SapI (1168) StuI (1292) BglII (1345) DrdI (1458) BciVI (1553) BseYI (1654) pGreenII 3304 bp
BsaBI  (3275)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
DraIII  (3087)
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  (3063)
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.
BspHI  (3031)
2 sites
T C A T G A A G T A C T
NruI  (2896)
1 site
T C G C G A A G C G C T
TsoI  (2714)
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  (2678)
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  (2640)
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.
SspI  (2628)
1 site
A A T A T T T T A T A A
AsiSI  (2555)
1 site
G C G A T C G C C G C T A G C G
Bpu10I  (2533)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
BsmBI  (2533)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
PflMI  (2293)
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.
BspHI  (2070)
2 sites
T C A T G A A G T A C T
AcuI  (1898)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (1766)
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  (1664)
1 site
G T G C A C C A C G T G
PspFI  (1658)
1 site
C C C A G C G G G T C G
StyI  (330)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NgoMIV  (433)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (435)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
FseI  (437)
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  (455)
1 site
T T A T A A A A T A T T
AanI  (455)
1 site
T T A T A A A A T A T T
PsiI  (455)
1 site
T T A T A A A A T A T T
BglII  (536)
2 sites
A G A T C T T C T A G A
HpaI  (568)
1 site
G T T A A C C A A T T G
FspI  (612)
1 site
T G C G C A A C G C G T
BmrI  (713)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
Acc65I  (783)
1 site
G G T A C C C C A T G G
KpnI  (787)
1 site
G G T A C C C C A T G G
PspOMI  (789)
1 site
G G G C C C C C C G G G
EcoO109I  (790)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
ApaI  (793)
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  (798)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (798)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (798)
1 site
V C T C G A G B B G A G C T C V
XhoI  (798)
1 site
C T C G A G G A G C T C
SalI  (804)
1 site
G T C G A C C A G C T G
AccI  (805)
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.
BspDI  (814)
1 site
A T C G A T T A G C T A
ClaI  (814)
1 site
A T C G A T T A G C T A
HindIII  (819)
1 site
A A G C T T T T C G A A
EcoRV  (827)
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  (831)
1 site
G A A T T C C T T A A G
PstI  (841)
1 site
C T G C A G G A C G T C
TspMI  (843)
1 site
C C C G G G G G G C C C
XmaI  (843)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (845)
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  (849)
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  (855)
1 site
A C T A G T T G A T C A
XbaI  (861)
1 site
T C T A G A A G A T C T
NotI  (868)
1 site
G C G G C C G C C G C C G G C G
BpmI  (873)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AleI  (879)
1 site
C A C N N N N G T G G T G N N N N C A C
MslI  (879)
1 site
C A Y N N N N R T G G T R N N N N Y A C
SacII  (880)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
BstXI  (881)
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.
Eco53kI  (887)
1 site
G A G C T C C T C G A G
SacI  (889)
1 site
G A G C T C C T C G A G
BspQI  (1168)
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  (1168)
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.
StuI  (1292)
1 site
A G G C C T T C C G G A
BglII  (1345)
2 sites
A G A T C T T C T A G A
DrdI  (1458)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BciVI  (1553)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
BseYI  (1654)
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.
KanR
2170 .. 2985  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
2170 .. 2985  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
lacZα
296 .. 946  =  651 bp
216 amino acids  =  23.7 kDa
Product: lacZα fragment of β-galactosidase
lacZα
296 .. 946  =  651 bp
216 amino acids  =  23.7 kDa
Product: lacZα fragment of β-galactosidase
ori
1411 .. 1999  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1411 .. 1999  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lac promoter
990 .. 1021  =  32 bp
   Segment 3:  -10  
   990 .. 996  =  7 bp
promoter for the E. coli lac operon
lac promoter
990 .. 1021  =  32 bp
   Segment 2:  
   997 .. 1015  =  19 bp
promoter for the E. coli lac operon
lac promoter
990 .. 1021  =  32 bp
   Segment 1:  -35  
   1016 .. 1021  =  6 bp
promoter for the E. coli lac operon
lac promoter
990 .. 1021  =  32 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
1296 .. 1320  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
1296 .. 1320  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
966 .. 982  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
966 .. 982  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
pSa ori
3276 .. 407  =  436 bp
origin of replication from bacterial plasmid pSa
pSa ori
3276 .. 407  =  436 bp
origin of replication from bacterial plasmid pSa
MCS
783 .. 890  =  108 bp
pBluescript multiple cloning site
MCS
783 .. 890  =  108 bp
pBluescript multiple cloning site
LB T-DNA repeat
542 .. 564  =  23 bp
left border repeat from nopaline C58 T-DNA
(truncated)
LB T-DNA repeat
542 .. 564  =  23 bp
left border repeat from nopaline C58 T-DNA
(truncated)
T7 promoter
756 .. 774  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
756 .. 774  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
903 .. 921  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
903 .. 921  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
733 .. 749  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
733 .. 749  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
942 .. 958  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
942 .. 958  =  17 bp
common sequencing primer, one of multiple similar
variants
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