pGreenII 0579

Agrobacterium binary vector with kanamycin- resistance and luciferase genes.

Sequence Author: pGreen website

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DraIII (5538) NruI (5347) PasI (5129) SspI (5079) AsiSI (5006) Bpu10I - BsmBI (4984) PflMI (4744) AlwNI (4217) ApaLI (4115) DrdI (3909) BglII (3796) StuI (3743) lac operator SacI (3340) Eco53kI (3338) SacII (3331) AleI - MslI (3330) NotI (3319) XbaI (3312) SpeI (3306) BamHI (3300) SmaI (3296) TspMI - XmaI (3294) PstI (3292) EcoRV (3279) HindIII (3271) AccI (3257) SalI (3256) AbsI - PaeR7I - PspXI - XhoI (3250) ApaI (3245) PspOMI (3241) KpnI (3239) Acc65I (3235) FspI (3064) BsaBI * (62) StyI (421) NgoMIV (524) NaeI (526) FseI (528) AanI (546) BglII (627) LB T-DNA repeat ScaI (677) BtgZI (918) SgrAI (1174) BfuAI - BspMI (1204) SexAI * (1262) PacI (1284) PpuMI (1424) XcmI (1872) NspI - SphI (1947) Bsu36I (1990) BstEII (1994) AflIII (2109) BsrGI (2112) AanI (2281) BsiWI (2448) PfoI * (2494) KasI (2570) NarI (2571) SfoI (2572) BbeI (2574) BmgBI (2738) PshAI (2801) pGreenII 0579 5664 bp
DraIII  (5538)
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.
NruI  (5347)
1 site
T C G C G A A G C G C T
PasI  (5129)
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.
SspI  (5079)
1 site
A A T A T T T T A T A A
AsiSI  (5006)
1 site
G C G A T C G C C G C T A G C G
Bpu10I  (4984)
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.
BsmBI  (4984)
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.
BsmBI-v2 is an improved version of BsmBI.
PflMI  (4744)
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  (4217)
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  (4115)
1 site
G T G C A C C A C G T G
DrdI  (3909)
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.
BglII  (3796)
2 sites
A G A T C T T C T A G A
StuI  (3743)
1 site
A G G C C T T C C G G A
SacI  (3340)
1 site
G A G C T C C T C G A G
Eco53kI  (3338)
1 site
G A G C T C C T C G A G
SacII  (3331)
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  (3330)
1 site
C A C N N N N G T G G T G N N N N C A C
MslI  (3330)
1 site
C A Y N N N N R T G G T R N N N N Y A C
NotI  (3319)
1 site
G C G G C C G C C G C C G G C G
XbaI  (3312)
1 site
T C T A G A A G A T C T
SpeI  (3306)
1 site
A C T A G T T G A T C A
BamHI  (3300)
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  (3296)
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  (3294)
1 site
C C C G G G G G G C C C
XmaI  (3294)
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.
PstI  (3292)
1 site
C T G C A G G A C G T C
EcoRV  (3279)
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  (3271)
1 site
A A G C T T T T C G A A
AccI  (3257)
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  (3256)
1 site
G T C G A C C A G C T G
AbsI  (3250)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (3250)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3250)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3250)
1 site
C T C G A G G A G C T C
ApaI  (3245)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (3241)
1 site
G G G C C C C C C G G G
KpnI  (3239)
1 site
G G T A C C C C A T G G
Acc65I  (3235)
1 site
G G T A C C C C A T G G
FspI  (3064)
1 site
T G C G C A A C G C G T
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)
2 sites
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
ScaI  (677)
1 site
A G T A C T T C A T G A
BtgZI  (918)
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.
SgrAI  (1174)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BfuAI  (1204)
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  (1204)
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.
SexAI  (1262)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
PacI  (1284)
1 site
T T A A T T A A A A T T A A T T
PpuMI  (1424)
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.
XcmI  (1872)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
NspI  (1947)
1 site
R C A T G Y Y G T A C R
SphI  (1947)
1 site
G C A T G C C G T A C G
Bsu36I  (1990)
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.
BstEII  (1994)
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.
AflIII  (2109)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
BsrGI  (2112)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
AanI  (2281)
2 sites
T T A T A A A A T A T T
BsiWI  (2448)
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.
PfoI  (2494)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
KasI  (2570)
1 site
G G C G C C C C G C G G
NarI  (2571)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2572)
1 site
G G C G C C C C G C G G
BbeI  (2574)
1 site
G G C G C C C C G C G G

Cleavage may be enhanced when more than one copy of the BbeI recognition sequence is present.
BmgBI  (2738)
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.
PshAI  (2801)
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.
luciferase
955 .. 2607  =  1653 bp
550 amino acids  =  60.7 kDa
Product: firefly luciferase
luciferase
955 .. 2607  =  1653 bp
550 amino acids  =  60.7 kDa
Product: firefly luciferase
KanR
4621 .. 5436  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
4621 .. 5436  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
3862 .. 4450  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3862 .. 4450  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
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 35S promoter
2618 .. 2963  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
2618 .. 2963  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
lacZα
3056 .. 3397  =  342 bp
113 amino acids  =  12.3 kDa
Product: LacZα fragment of β-galactosidase
lacZα
3056 .. 3397  =  342 bp
113 amino acids  =  12.3 kDa
Product: LacZα fragment of β-galactosidase
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
3441 .. 3472  =  32 bp
3 segments
   Segment 3:  -10  
   3441 .. 3447  =  7 bp
promoter for the E. coli lac operon
lac promoter
3441 .. 3472  =  32 bp
3 segments
   Segment 2:  
   3448 .. 3466  =  19 bp
promoter for the E. coli lac operon
lac promoter
3441 .. 3472  =  32 bp
3 segments
   Segment 1:  -35  
   3467 .. 3472  =  6 bp
promoter for the E. coli lac operon
lac promoter
3441 .. 3472  =  32 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
3747 .. 3771  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
3747 .. 3771  =  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
3417 .. 3433  =  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
3417 .. 3433  =  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).
MCS
3235 .. 3341  =  107 bp
pBluescript multiple cloning site
MCS
3235 .. 3341  =  107 bp
pBluescript multiple cloning site
T7 promoter
3208 .. 3226  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3208 .. 3226  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
3354 .. 3372  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
3354 .. 3372  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
3185 .. 3201  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
3185 .. 3201  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
3393 .. 3409  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
3393 .. 3409  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  3783 .. 4187  =  405 bp
ORF:  134 amino acids  =  15.0 kDa
ORF:  4599 .. 4826  =  228 bp
ORF:  75 amino acids  =  8.8 kDa
ORF:  955 .. 2691  =  1737 bp
ORF:  578 amino acids  =  64.1 kDa
ORF:  4621 .. 5436  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  2717 .. 3025  =  309 bp
ORF:  102 amino acids  =  11.3 kDa
ORF:  3056 .. 3397  =  342 bp
ORF:  113 amino acids  =  12.3 kDa
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