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Plasmid Files

pGreenII 0179

Agrobacterium binary vector with kanamycin- and hygromycin-resistance genes.

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pGreenII 0179 Sequence and MappGreenII 0179.dna
Map and Sequence File   
Sequence Author:  pGreen website
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 NruI (4821) PasI (4603) EcoNI (4565) SspI (4553) PflMI (4218) AlwNI (3691) BciVI (3478) BglII (3270) StuI (3217) BspQI - SapI (3093) lac operator SacI (2814) Eco53kI (2812) BstXI (2806) SacII (2805) AleI - MslI (2804) NotI (2793) XbaI (2786) SpeI (2780) BamHI (2774) SmaI (2770) TspMI - XmaI (2768) PstI (2766) EcoRI (2756) EcoRV (2752) HindIII (2744) AccI (2730) SalI (2729) AbsI - PaeR7I - PspXI - XhoI (2723) ApaI (2718) EcoO109I (2715) PspOMI (2714) KpnI (2712) Acc65I (2708) BsaBI * (62) StyI (421) NgoMIV (524) NaeI (526) FseI (528) AanI - PsiI (546) BglII (627) LB T-DNA repeat BlpI (919) BbvCI (1005) BtgZI (1397) NdeI (1621) RsrII (1662) BfuAI - BspMI (1756) ZraI (2048) AatII (2050) BmgBI (2211) FspI (2537) BmrI (2638) pGreenII 0179 5138 bp
NruI  (4821)
1 site
T C G C G A A G C G C T
PasI  (4603)
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  (4565)
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  (4553)
1 site
A A T A T T T T A T A A
PflMI  (4218)
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  (3691)
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.
BciVI  (3478)
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.
BglII  (3270)
2 sites
A G A T C T T C T A G A
StuI  (3217)
1 site
A G G C C T T C C G G A
BspQI  (3093)
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  (3093)
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  (2814)
1 site
G A G C T C C T C G A G
Eco53kI  (2812)
1 site
G A G C T C C T C G A G
BstXI  (2806)
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.
SacII  (2805)
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.
AleI  (2804)
1 site
C A C N N N N G T G G T G N N N N C A C
MslI  (2804)
1 site
C A Y N N N N R T G G T R N N N N Y A C
NotI  (2793)
1 site
G C G G C C G C C G C C G G C G
XbaI  (2786)
1 site
T C T A G A A G A T C T
SpeI  (2780)
1 site
A C T A G T T G A T C A
BamHI  (2774)
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.
SmaI  (2770)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (2768)
1 site
C C C G G G G G G C C C
XmaI  (2768)
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.
PstI  (2766)
1 site
C T G C A G G A C G T C
EcoRI  (2756)
1 site
G A A T T C C T T A A G
EcoRV  (2752)
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.
HindIII  (2744)
1 site
A A G C T T T T C G A A
AccI  (2730)
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.
SalI  (2729)
1 site
G T C G A C C A G C T G
AbsI  (2723)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2723)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2723)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2723)
1 site
C T C G A G G A G C T C
ApaI  (2718)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoO109I  (2715)
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.
PspOMI  (2714)
1 site
G G G C C C C C C G G G
KpnI  (2712)
1 site
G G T A C C C C A T G G
Acc65I  (2708)
1 site
G G T A C C C C A T G G
BsaBI  (62)
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.
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.
NgoMIV  (524)
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  (526)
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  (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
BlpI  (919)
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.
BbvCI  (1005)
1 site
C C T C A G C G G A G T C G
BtgZI  (1397)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
NdeI  (1621)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
RsrII  (1662)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BfuAI  (1756)
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  (1756)
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.
ZraI  (2048)
1 site
G A C G T C C T G C A G
AatII  (2050)
1 site
G A C G T C C T G C A G
BmgBI  (2211)
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.
FspI  (2537)
1 site
T G C G C A A C G C G T
BmrI  (2638)
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.
HygR
1049 .. 2074  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
1049 .. 2074  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
KanR
4095 .. 4910  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
4095 .. 4910  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
ori
3336 .. 3924  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3336 .. 3924  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lacZα
2434 .. 2871  =  438 bp
145 amino acids  =  16.2 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2434 .. 2871  =  438 bp
145 amino acids  =  16.2 kDa
Product: LacZα fragment of β-galactosidase
pSa ori
63 .. 498  =  436 bp
origin of replication from bacterial plasmid pSa
pSa ori
63 .. 498  =  436 bp
origin of replication from bacterial plasmid pSa
CaMV poly(A) signal
663 .. 839  =  177 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
663 .. 839  =  177 bp
cauliflower mosaic virus polyadenylation signal
lac promoter
2915 .. 2946  =  32 bp
   Segment 3:  -10  
   2915 .. 2921  =  7 bp
promoter for the E. coli lac operon
lac promoter
2915 .. 2946  =  32 bp
   Segment 2:  
   2922 .. 2940  =  19 bp
promoter for the E. coli lac operon
lac promoter
2915 .. 2946  =  32 bp
   Segment 1:  -35  
   2941 .. 2946  =  6 bp
promoter for the E. coli lac operon
lac promoter
2915 .. 2946  =  32 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
3221 .. 3245  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
3221 .. 3245  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
633 .. 655  =  23 bp
left border repeat from nopaline C58 T-DNA
(truncated)
LB T-DNA repeat
633 .. 655  =  23 bp
left border repeat from nopaline C58 T-DNA
(truncated)
lac operator
2891 .. 2907  =  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
2891 .. 2907  =  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).
CaMV 35S promoter
2091 .. 2436  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
2091 .. 2436  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
MCS
2708 .. 2815  =  108 bp
pBluescript multiple cloning site
MCS
2708 .. 2815  =  108 bp
pBluescript multiple cloning site
T7 promoter
2681 .. 2699  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2681 .. 2699  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
2828 .. 2846  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
2828 .. 2846  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
2658 .. 2674  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
2658 .. 2674  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2867 .. 2883  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2867 .. 2883  =  17 bp
common sequencing primer, one of multiple similar
variants
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