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Plasmid Files

pPZP221

Agrobacterium binary vector for plant transformation, with spectinomycin- and gentamycin-resistance genes. The MCS is reversed in pPZP222.

 
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 PmeI (8580) HindIII (8367) PstI - SbfI (8359) SalI (8349) XbaI (8343) BamHI (8337) SmaI (8334) KpnI - TspMI - XmaI (8332) Acc65I (8328) BanII - SacI (8326) Eco53kI (8324) EcoRI (8316) lac operator BstXI (8073) NcoI (7260) PmlI (6627) BstEII (5707) BpmI (5622) BlpI (5560) BssHII (5144) HpaI (5033) NsiI (4913) PasI (1306) AclI (1900) BsiWI (2860) NheI (2976) BmtI (2980) BspDI * - ClaI * (3056) EcoNI (3289) BsaI (3379) AgeI (3470) KasI (3761) NarI (3762) SfoI (3763) PluTI (3765) BstZ17I (3960) pPZP221 8732 bp
PmeI  (8580)
1 site
G T T T A A A C C A A A T T T G
HindIII  (8367)
1 site
A A G C T T T T C G A A
PstI  (8359)
1 site
C T G C A G G A C G T C
SbfI  (8359)
1 site
C C T G C A G G G G A C G T C C
SalI  (8349)
1 site
G T C G A C C A G C T G
XbaI  (8343)
1 site
T C T A G A A G A T C T
BamHI  (8337)
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  (8334)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KpnI  (8332)
1 site
G G T A C C C C A T G G
TspMI  (8332)
1 site
C C C G G G G G G C C C
XmaI  (8332)
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.
Acc65I  (8328)
1 site
G G T A C C C C A T G G
BanII  (8326)
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  (8326)
1 site
G A G C T C C T C G A G
Eco53kI  (8324)
1 site
G A G C T C C T C G A G
EcoRI  (8316)
1 site
G A A T T C C T T A A G
BstXI  (8073)
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.
NcoI  (7260)
1 site
C C A T G G G G T A C C
PmlI  (6627)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstEII  (5707)
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.
BpmI  (5622)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BlpI  (5560)
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.
BssHII  (5144)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
HpaI  (5033)
1 site
G T T A A C C A A T T G
NsiI  (4913)
1 site
A T G C A T T A C G T A
PasI  (1306)
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.
AclI  (1900)
1 site
A A C G T T T T G C A A
BsiWI  (2860)
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.
NheI  (2976)
1 site
G C T A G C C G A T C G
BmtI  (2980)
1 site
G C T A G C C G A T C G
BspDI  (3056)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3056)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoNI  (3289)
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.
BsaI  (3379)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AgeI  (3470)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
KasI  (3761)
1 site
G G C G C C C C G C G G
NarI  (3762)
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  (3763)
1 site
G G C G C C C C G C G G
PluTI  (3765)
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.
BstZ17I  (3960)
1 site
G T A T A C C A T A T G
pVS1 RepA
2245 .. 3318  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
2245 .. 3318  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
SmR
5080 .. 5871  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and
streptomycin
SmR
5080 .. 5871  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and
streptomycin
CaMV 35S promoter (enhanced)
7323 .. 8001  =  679 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
7323 .. 8001  =  679 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
1187 .. 1816  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
1187 .. 1816  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
4248 .. 4836  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4248 .. 4836  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
GmR
6730 .. 7263  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
6730 .. 7263  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
lacZα
8302 .. 8535  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8302 .. 8535  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
3384 .. 3578  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
3384 .. 3578  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
6521 .. 6695  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6521 .. 6695  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
3922 .. 4062  =  141 bp
basis of mobility region from pBR322
bom
3922 .. 4062  =  141 bp
basis of mobility region from pBR322
lac promoter
8228 .. 8258  =  31 bp
   Segment 1:  -35  
   8228 .. 8233  =  6 bp
promoter for the E. coli lac operon
lac promoter
8228 .. 8258  =  31 bp
   Segment 2:  
   8234 .. 8251  =  18 bp
promoter for the E. coli lac operon
lac promoter
8228 .. 8258  =  31 bp
   Segment 3:  -10  
   8252 .. 8258  =  7 bp
promoter for the E. coli lac operon
lac promoter
8228 .. 8258  =  31 bp
3 segments
promoter for the E. coli lac operon
LB T-DNA repeat
6399 .. 6423  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6399 .. 6423  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8595 .. 8619  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8595 .. 8619  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
8266 .. 8282  =  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
8266 .. 8282  =  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
8316 .. 8372  =  57 bp
pUC18 multiple cloning site
MCS
8316 .. 8372  =  57 bp
pUC18 multiple cloning site
M13 rev
8290 .. 8306  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
8290 .. 8306  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
8376 .. 8392  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
8376 .. 8392  =  17 bp
common sequencing primer, one of multiple similar
variants
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