pFGC5941

Agrobacterium binary vector for expressing dsRNA, with two multiple cloning sites separated by an intron.
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BclI * (11,169) EcoRV (10,289) BstZ17I (9225) PluTI (9030) SfoI (9028) NarI (9027) KasI (9026) EcoNI (8554) BsiWI (8125) LB T-DNA repeat Acc65I (407) KpnI (411) FspAI (521) PaqCI (585) SalI (656) AfeI (758) EcoRI (1317) MluI (1536) StuI (1867) PshAI (2466) PaeR7I - PspXI - XhoI (2653) TMV Ω NcoI (2734) AscI (2741) SwaI (2760) AflII (3659) PmlI (3675) BamHI (4122) AvrII (4126) XbaI (4136) PacI (4149) TspMI - XmaI (4155) SmaI (4157) MfeI (4465) BstEII (4737) AhdI (4867) SbfI (4892) PvuI (5026) PmeI (5113) pFGC5941 11,406 bp
BclI  (11,169)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
EcoRV  (10,289)
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.
BstZ17I  (9225)
1 site
G T A T A C C A T A T G
PluTI  (9030)
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.
SfoI  (9028)
1 site
G G C G C C C C G C G G
NarI  (9027)
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.
KasI  (9026)
1 site
G G C G C C C C G C G G
EcoNI  (8554)
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.
BsiWI  (8125)
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.
Acc65I  (407)
1 site
G G T A C C C C A T G G
KpnI  (411)
1 site
G G T A C C C C A T G G
FspAI  (521)
1 site
R T G C G C A Y Y A C G C G T R
PaqCI  (585)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
SalI  (656)
1 site
G T C G A C C A G C T G
AfeI  (758)
1 site
A G C G C T T C G C G A
EcoRI  (1317)
1 site
G A A T T C C T T A A G
MluI  (1536)
1 site
A C G C G T T G C G C A
StuI  (1867)
1 site
A G G C C T T C C G G A
PshAI  (2466)
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.
PaeR7I  (2653)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2653)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2653)
1 site
C T C G A G G A G C T C
NcoI  (2734)
1 site
C C A T G G G G T A C C
AscI  (2741)
1 site
G G C G C G C C C C G C G C G G
SwaI  (2760)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AflII  (3659)
1 site
C T T A A G G A A T T C
PmlI  (3675)
1 site
C A C G T G G T G C A C
BamHI  (4122)
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.
AvrII  (4126)
1 site
C C T A G G G G A T C C
XbaI  (4136)
1 site
T C T A G A A G A T C T
PacI  (4149)
1 site
T T A A T T A A A A T T A A T T
TspMI  (4155)
1 site
C C C G G G G G G C C C
XmaI  (4155)
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  (4157)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
MfeI  (4465)
1 site
C A A T T G G T T A A C
BstEII  (4737)
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.
AhdI  (4867)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
SbfI  (4892)
1 site
C C T G C A G G G G A C G T C C
PvuI  (5026)
1 site
C G A T C G G C T A G C
PmeI  (5113)
1 site
G T T T A A A C C A A A T T T G
chsA intron
2767 .. 4115  =  1349 bp
chalcone synthase A intron from Petunia hybrida
chsA intron
2767 .. 4115  =  1349 bp
chalcone synthase A intron from Petunia hybrida
pVS1 RepA
7510 .. 8583  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
pVS1 RepA
7510 .. 8583  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas plasmid pVS1
KanR
10,188 .. 10,982  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
10,188 .. 10,982  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
OCS terminator
4177 .. 4884  =  708 bp
octopine synthase terminator
OCS terminator
4177 .. 4884  =  708 bp
octopine synthase terminator
pVS1 StaA
6452 .. 7081  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas plasmid pVS1
pVS1 StaA
6452 .. 7081  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas plasmid pVS1
ori
9513 .. 10,101  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9513 .. 10,101  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
BlpR
373 .. 924  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
BlpR
373 .. 924  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
MAS promoter
930 .. 1310  =  381 bp
mannopine synthase promoter (Velten et al., 1984)
MAS promoter
930 .. 1310  =  381 bp
mannopine synthase promoter (Velten et al., 1984)
CaMV 35S promoter
2305 .. 2650  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
2305 .. 2650  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
MAS terminator
111 .. 363  =  253 bp
mannopine synthase terminator
MAS terminator
111 .. 363  =  253 bp
mannopine synthase terminator
pVS1 oriV
8649 .. 8843  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
pVS1 oriV
8649 .. 8843  =  195 bp
origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)
bom
9187 .. 9327  =  141 bp
basis of mobility region from pBR322
bom
9187 .. 9327  =  141 bp
basis of mobility region from pBR322
TMV Ω
2680 .. 2734  =  55 bp
translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)
TMV Ω
2680 .. 2734  =  55 bp
translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)
MCS 2
4122 .. 4160  =  39 bp
multiple cloning site
MCS 2
4122 .. 4160  =  39 bp
multiple cloning site
LB T-DNA repeat
1 .. 25  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
1 .. 25  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
5128 .. 5152  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
5128 .. 5152  =  25 bp
right border repeat from nopaline C58 T-DNA
MCS 1
2734 .. 2764  =  31 bp
multiple cloning site 1
MCS 1
2734 .. 2764  =  31 bp
multiple cloning site 1
ORF:  448 .. 957  =  510 bp
ORF:  169 amino acids  =  18.0 kDa
ORF:  1729 .. 2133  =  405 bp
ORF:  134 amino acids  =  15.5 kDa
ORF:  5467 .. 6153  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  7408 .. 8583  =  1176 bp
ORF:  391 amino acids  =  43.9 kDa
ORF:  10,321 .. 10,551  =  231 bp
ORF:  76 amino acids  =  8.3 kDa
ORF:  1808 .. 2071  =  264 bp
ORF:  87 amino acids  =  10.4 kDa
ORF:  2123 .. 2551  =  429 bp
ORF:  142 amino acids  =  15.9 kDa
ORF:  6452 .. 7081  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  4764 .. 5054  =  291 bp
ORF:  96 amino acids  =  10.1 kDa
ORF:  11,227 .. 321  =  501 bp
ORF:  166 amino acids  =  18.5 kDa
ORF:  373 .. 924  =  552 bp
ORF:  183 amino acids  =  20.6 kDa
ORF:  1756 .. 2124  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  6190 .. 6504  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  7378 .. 7950  =  573 bp
ORF:  190 amino acids  =  20.6 kDa
ORF:  5499 .. 5849  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  7356 .. 7613  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  10,188 .. 10,982  =  795 bp
ORF:  264 amino acids  =  31.0 kDa
ORF:  5918 .. 6190  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
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