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Plasmid Files

pGreen 0029

Compact Agrobacterium binary vector with a kanamycin-resistance genes for bacterial and plant transformation.

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pGreen 0029 Sequence and MappGreen 0029.dna
Map and Sequence File   
Sequence Author:  pGreen website
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 RsrII (4505) AflII (4285) AflIII - PciI (4205) MauBI (4091) SphI (4061) PsiI (3937) FseI (3919) StyI (3812) DraIII (3265) BspHI (3209) NruI (3074) EcoNI (2818) AsiSI (2733) Bpu10I - BsmBI (2711) PflMI (2471) NmeAIII (170) PflFI - Tth111I (273) MscI (310) KasI (389) BsaHI - NarI (390) SfoI (391) PluTI (393) NheI (616) BmtI (620) Bsu36I (632) BlpI (677) Acc65I (1031) KpnI (1035) PspOMI (1037) EcoO109I (1038) ApaI (1041) AbsI - PaeR7I - PspXI - XhoI (1046) SalI (1052) AccI (1053) HincII (1054) HindIII (1067) EcoRV (1075) EcoRI (1079) PstI (1089) TspMI - XmaI (1091) SmaI (1093) BamHI (1097) SpeI (1103) XbaI (1109) NotI (1116) AleI - MslI (1127) BstXI (1129) Eco53kI (1135) SacI (1137) lac operator StuI (1540) ApaLI (1912) AlwNI (2014) pGreen 0029 4632 bp
RsrII  (4505)
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.
AflII  (4285)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
AflIII  (4205)
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  (4205)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
MauBI  (4091)
1 site
C G C G C G C G G C G C G C G C
SphI  (4061)
1 site
G C A T G C C G T A C G
PsiI  (3937)
1 site
T T A T A A A A T A T T
FseI  (3919)
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.
StyI  (3812)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
DraIII  (3265)
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.
BspHI  (3209)
1 site
T C A T G A A G T A C T
NruI  (3074)
1 site
T C G C G A A G C G C T
EcoNI  (2818)
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  (2733)
1 site
G C G A T C G C C G C T A G C G
Bpu10I  (2711)
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  (2711)
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  (2471)
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.
NmeAIII  (170)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PflFI  (273)
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  (273)
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.
MscI  (310)
1 site
T G G C C A A C C G G T
KasI  (389)
1 site
G G C G C C C C G C G G
BsaHI  (390)
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.
NarI  (390)
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  (391)
1 site
G G C G C C C C G C G G
PluTI  (393)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
NheI  (616)
1 site
G C T A G C C G A T C G
BmtI  (620)
1 site
G C T A G C C G A T C G
Bsu36I  (632)
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  (677)
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  (1031)
1 site
G G T A C C C C A T G G
KpnI  (1035)
1 site
G G T A C C C C A T G G
PspOMI  (1037)
1 site
G G G C C C C C C G G G
EcoO109I  (1038)
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  (1041)
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  (1046)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (1046)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1046)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1046)
1 site
C T C G A G G A G C T C
SalI  (1052)
1 site
G T C G A C C A G C T G
AccI  (1053)
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.
HincII  (1054)
1 site
G T Y R A C C A R Y T G
HindIII  (1067)
1 site
A A G C T T T T C G A A
EcoRV  (1075)
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  (1079)
1 site
G A A T T C C T T A A G
PstI  (1089)
1 site
C T G C A G G A C G T C
TspMI  (1091)
1 site
C C C G G G G G G C C C
XmaI  (1091)
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  (1093)
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  (1097)
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  (1103)
1 site
A C T A G T T G A T C A
XbaI  (1109)
1 site
T C T A G A A G A T C T
NotI  (1116)
1 site
G C G G C C G C C G C C G G C G
AleI  (1127)
1 site
C A C N N N N G T G G T G N N N N C A C
MslI  (1127)
1 site
C A Y N N N N R T G G T R N N N N Y A C
BstXI  (1129)
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  (1135)
1 site
G A G C T C C T C G A G
SacI  (1137)
1 site
G A G C T C C T C G A G
StuI  (1540)
1 site
A G G C C T T C C G G A
ApaLI  (1912)
1 site
G T G C A C C A C G T G
AlwNI  (2014)
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.
KanR
2348 .. 3163  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
2348 .. 3163  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
NeoR/KanR
4359 .. 521  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
4359 .. 521  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
1659 .. 2247  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1659 .. 2247  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
pSa ori
3454 .. 3889  =  436 bp
origin of replication from bacterial plasmid pSa
pSa ori
3454 .. 3889  =  436 bp
origin of replication from bacterial plasmid pSa
lacZα
805 .. 1194  =  390 bp
129 amino acids  =  14.2 kDa
Product: LacZα fragment of β-galactosidase
lacZα
805 .. 1194  =  390 bp
129 amino acids  =  14.2 kDa
Product: LacZα fragment of β-galactosidase
NOS terminator
4068 .. 4319  =  252 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
4068 .. 4319  =  252 bp
nopaline synthase terminator and poly(A) signal
NOS promoter
555 .. 734  =  180 bp
nopaline synthase promoter
NOS promoter
555 .. 734  =  180 bp
nopaline synthase promoter
lac promoter
1238 .. 1269  =  32 bp
   Segment 3:  -10  
   1238 .. 1244  =  7 bp
promoter for the E. coli lac operon
lac promoter
1238 .. 1269  =  32 bp
   Segment 2:  
   1245 .. 1263  =  19 bp
promoter for the E. coli lac operon
lac promoter
1238 .. 1269  =  32 bp
   Segment 1:  -35  
   1264 .. 1269  =  6 bp
promoter for the E. coli lac operon
lac promoter
1238 .. 1269  =  32 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
1544 .. 1568  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
1544 .. 1568  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
4022 .. 4044  =  23 bp
left border repeat from nopaline C58 T-DNA
(truncated)
LB T-DNA repeat
4022 .. 4044  =  23 bp
left border repeat from nopaline C58 T-DNA
(truncated)
lac operator
1214 .. 1230  =  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
1214 .. 1230  =  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
1031 .. 1138  =  108 bp
pBluescript multiple cloning site
MCS
1031 .. 1138  =  108 bp
pBluescript multiple cloning site
T7 promoter
1004 .. 1022  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1004 .. 1022  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
1151 .. 1169  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
1151 .. 1169  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
981 .. 997  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
981 .. 997  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
1190 .. 1206  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
1190 .. 1206  =  17 bp
common sequencing primer, one of multiple similar
variants
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