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Plasmid Files

pSoup

Plant vector used for co-transformation with pGreen family plasmids.

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pSoup Sequence and MappSoup.dna
Map and Sequence File   
Sequence Author:  pGreen website
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 BclI * (9210) RsrII (9105) AatII (8338) ZraI (8336) PmlI (7830) AgeI (7762) MfeI (5975) AleI (5705) NdeI (5579) PfoI * (5487) SfiI (5021) PciI (235) DrdI (343) NotI - SacII (1367) MscI (1514) PsiI (1806) PasI (1920) Eco53kI (2450) SacI (2452) Acc65I (2454) KpnI - TspMI - XmaI (2458) SmaI (2460) BamHI (2463) XbaI (2469) SalI (2475) PstI - SbfI (2485) SphI (2491) HindIII (2493) SP6 promoter lac operator PvuII (2706) XmnI (2890) tet promoter EcoRV (3079) NheI (3121) BmtI (3125) PshAI (3608) pSoup 9275 bp
BclI  (9210)
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.
RsrII  (9105)
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.
AatII  (8338)
1 site
G A C G T C C T G C A G
ZraI  (8336)
1 site
G A C G T C C T G C A G
PmlI  (7830)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AgeI  (7762)
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.
MfeI  (5975)
1 site
C A A T T G G T T A A C
AleI  (5705)
1 site
C A C N N N N G T G G T G N N N N C A C
NdeI  (5579)
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.
PfoI  (5487)
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.
SfiI  (5021)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
PciI  (235)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (343)
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.
NotI  (1367)
1 site
G C G G C C G C C G C C G G C G
SacII  (1367)
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.
MscI  (1514)
1 site
T G G C C A A C C G G T
PsiI  (1806)
1 site
T T A T A A A A T A T T
PasI  (1920)
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.
Eco53kI  (2450)
1 site
G A G C T C C T C G A G
SacI  (2452)
1 site
G A G C T C C T C G A G
Acc65I  (2454)
1 site
G G T A C C C C A T G G
KpnI  (2458)
1 site
G G T A C C C C A T G G
TspMI  (2458)
1 site
C C C G G G G G G C C C
XmaI  (2458)
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  (2460)
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  (2463)
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.
XbaI  (2469)
1 site
T C T A G A A G A T C T
SalI  (2475)
1 site
G T C G A C C A G C T G
PstI  (2485)
1 site
C T G C A G G A C G T C
SbfI  (2485)
1 site
C C T G C A G G G G A C G T C C
SphI  (2491)
1 site
G C A T G C C G T A C G
HindIII  (2493)
1 site
A A G C T T T T C G A A
PvuII  (2706)
1 site
C A G C T G G T C G A C
XmnI  (2890)
1 site
G A A N N N N T T C C T T N N N N A A G
EcoRV  (3079)
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.
NheI  (3121)
1 site
G C T A G C C G A T C G
BmtI  (3125)
1 site
G C T A G C C G A T C G
PshAI  (3608)
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.
TcR
2978 .. 4168  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
2978 .. 4168  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
trfA
4667 .. 5815  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds
to and activates oriV
trfA
4667 .. 5815  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds
to and activates oriV
RepA
7196 .. 8167  =  972 bp
323 amino acids  =  36.5 kDa
Product: replication protein of plasmid pSa
RepA
7196 .. 8167  =  972 bp
323 amino acids  =  36.5 kDa
Product: replication protein of plasmid pSa
oriV
1258 .. 1969  =  712 bp
incP origin of replication
oriV
1258 .. 1969  =  712 bp
incP origin of replication
TetR
8562 .. 9212  =  651 bp
216 amino acids  =  23.3 kDa
Product: tetracycline resistance regulatory protein
TetR
8562 .. 9212  =  651 bp
216 amino acids  =  23.3 kDa
Product: tetracycline resistance regulatory protein
traJ
760 .. 1131  =  372 bp
123 amino acids  =  13.5 kDa
Product: oriT-recognizing protein
traJ
760 .. 1131  =  372 bp
123 amino acids  =  13.5 kDa
Product: oriT-recognizing protein
oriT
618 .. 727  =  110 bp
incP origin of transfer
oriT
618 .. 727  =  110 bp
incP origin of transfer
MCS
2448 .. 2498  =  51 bp
multiple cloning site
MCS
2448 .. 2498  =  51 bp
multiple cloning site
lac promoter
2589 .. 2619  =  31 bp
   Segment 3:  -10  
   2589 .. 2595  =  7 bp
promoter for the E. coli lac operon
lac promoter
2589 .. 2619  =  31 bp
   Segment 2:  
   2596 .. 2613  =  18 bp
promoter for the E. coli lac operon
lac promoter
2589 .. 2619  =  31 bp
   Segment 1:  -35  
   2614 .. 2619  =  6 bp
promoter for the E. coli lac operon
lac promoter
2589 .. 2619  =  31 bp
3 segments
promoter for the E. coli lac operon
tet promoter
2902 .. 2930  =  29 bp
   Segment 1:  -35  
   2902 .. 2907  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
2902 .. 2930  =  29 bp
   Segment 2:  
   2908 .. 2924  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
2902 .. 2930  =  29 bp
   Segment 3:  -10  
   2925 .. 2930  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
2902 .. 2930  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
SP6 promoter
2505 .. 2523  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
2505 .. 2523  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 rev
2541 .. 2557  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2541 .. 2557  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
2565 .. 2581  =  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
2565 .. 2581  =  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).
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