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Plasmid Files

pBI221

Transient expression vector with a GUS reporter gene for plant transformation.

 
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 HindIII (1) lac operator BspQI - SapI (5426) AhdI (4421) NmeAIII (4274) AatII (3502) ZraI (3500) BstAPI (3251) NdeI (3247) PluTI (3198) SfoI (3196) NarI (3195) KasI (3194) EcoRI (3033) MauBI (3002) NsiI (2898) SphI (11) PstI - SbfI (17) NcoI (328) AccI (463) BspEI (537) BmgBI (736) XbaI (866) BamHI (872) TspMI - XmaI (877) SmaI (879) BclI * (985) BsaAI - SnaBI (1280) MscI (1517) BstBI (1989) BsgI (2417) Eco53kI (2764) BanII - SacI (2766) AflII (2807) pBI221 5667 bp
HindIII  (1)
1 site
A A G C T T T T C G A A
BspQI  (5426)
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  (5426)
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.
AhdI  (4421)
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.
NmeAIII  (4274)
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).
AatII  (3502)
1 site
G A C G T C C T G C A G
ZraI  (3500)
1 site
G A C G T C C T G C A G
BstAPI  (3251)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
NdeI  (3247)
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.
PluTI  (3198)
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  (3196)
1 site
G G C G C C C C G C G G
NarI  (3195)
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  (3194)
1 site
G G C G C C C C G C G G
EcoRI  (3033)
1 site
G A A T T C C T T A A G
MauBI  (3002)
1 site
C G C G C G C G G C G C G C G C
NsiI  (2898)
1 site
A T G C A T T A C G T A
SphI  (11)
1 site
G C A T G C C G T A C G
PstI  (17)
1 site
C T G C A G G A C G T C
SbfI  (17)
1 site
C C T G C A G G G G A C G T C C
NcoI  (328)
1 site
C C A T G G G G T A C C
AccI  (463)
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.
BspEI  (537)
1 site
T C C G G A A G G C C T
BmgBI  (736)
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.
XbaI  (866)
1 site
T C T A G A A G A T C T
BamHI  (872)
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.
TspMI  (877)
1 site
C C C G G G G G G C C C
XmaI  (877)
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  (879)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BclI  (985)
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.
BsaAI  (1280)
1 site
Y A C G T R R T G C A Y
SnaBI  (1280)
1 site
T A C G T A A T G C A T
MscI  (1517)
1 site
T G G C C A A C C G G T
BstBI  (1989)
1 site
T T C G A A A A G C T T
BsgI  (2417)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
Eco53kI  (2764)
1 site
G A G C T C C T C G A G
BanII  (2766)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (2766)
1 site
G A G C T C C T C G A G
AflII  (2807)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
GUS
896 .. 2707  =  1812 bp
603 amino acids  =  68.4 kDa
Product: β-glucuronidase
GUS
896 .. 2707  =  1812 bp
603 amino acids  =  68.4 kDa
Product: β-glucuronidase
AmpR
3634 .. 4494  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3634 .. 3702  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3634 .. 4494  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3703 .. 4494  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3634 .. 4494  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
4665 .. 5253  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4665 .. 5253  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CaMV 35S promoter
512 .. 857  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
512 .. 857  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
2778 .. 3030  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
2778 .. 3030  =  253 bp
nopaline synthase terminator and poly(A) signal
AmpR promoter
3529 .. 3633  =  105 bp
AmpR promoter
3529 .. 3633  =  105 bp
lac promoter
5577 .. 5607  =  31 bp
   Segment 1:  -35  
   5577 .. 5582  =  6 bp
promoter for the E. coli lac operon
lac promoter
5577 .. 5607  =  31 bp
   Segment 2:  
   5583 .. 5600  =  18 bp
promoter for the E. coli lac operon
lac promoter
5577 .. 5607  =  31 bp
   Segment 3:  -10  
   5601 .. 5607  =  7 bp
promoter for the E. coli lac operon
lac promoter
5577 .. 5607  =  31 bp
3 segments
promoter for the E. coli lac operon
M13 fwd
3039 .. 3055  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
3039 .. 3055  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
5615 .. 5631  =  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
5615 .. 5631  =  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).
M13 rev
5639 .. 5655  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
5639 .. 5655  =  17 bp
common sequencing primer, one of multiple similar
variants
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