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Plasmid Files

pRS313

Yeast centromere vector with a HIS3 marker and an MCS derived from pBLUESCRIPT.

 
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 SwaI (4423) PmlI (4399) PpuMI (4385) BsaHI (4003) ScaI (3946) BpmI (3536) BsaI (3527) AhdI (3466) AlwNI (2989) PfoI (46) AfeI (211) BseRI (301) BsaBI * (560) MscI (710) BsmI (715) BsiWI (915) NheI (1007) BmtI (1011) BclI * (1051) NsiI (1282) BssHII (1311) DraIII (1647) BtgZI (1648) NgoMIV (1748) NaeI (1750) Eco53kI (2075) SacI (2077) BtgI (2081) SacII (2084) EagI - NotI (2090) XbaI (2097) SpeI (2103) BamHI (2109) TspMI - XmaI (2115) SmaI (2117) EcoRI (2127) EcoRV (2135) BspDI - ClaI (2146) SalI (2154) HincII (2156) AbsI - PaeR7I - PspXI - XhoI (2160) PspOMI (2169) ApaI (2173) lac operator pRS313 4967 bp
SwaI  (4423)
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.
PmlI  (4399)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
PpuMI  (4385)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BsaHI  (4003)
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.
ScaI  (3946)
1 site
A G T A C T T C A T G A
BpmI  (3536)
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.
BsaI  (3527)
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.
AhdI  (3466)
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.
AlwNI  (2989)
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.
PfoI  (46)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
AfeI  (211)
1 site
A G C G C T T C G C G A
BseRI  (301)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
BsaBI  (560)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MscI  (710)
1 site
T G G C C A A C C G G T
BsmI  (715)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BsiWI  (915)
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  (1007)
1 site
G C T A G C C G A T C G
BmtI  (1011)
1 site
G C T A G C C G A T C G
BclI  (1051)
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.
NsiI  (1282)
1 site
A T G C A T T A C G T A
BssHII  (1311)
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.
DraIII  (1647)
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.
BtgZI  (1648)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
NgoMIV  (1748)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (1750)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
Eco53kI  (2075)
1 site
G A G C T C C T C G A G
SacI  (2077)
1 site
G A G C T C C T C G A G
BtgI  (2081)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (2084)
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.
EagI  (2090)
1 site
C G G C C G G C C G G C
NotI  (2090)
1 site
G C G G C C G C C G C C G G C G
XbaI  (2097)
1 site
T C T A G A A G A T C T
SpeI  (2103)
1 site
A C T A G T T G A T C A
BamHI  (2109)
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  (2115)
1 site
C C C G G G G G G C C C
XmaI  (2115)
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  (2117)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (2127)
1 site
G A A T T C C T T A A G
EcoRV  (2135)
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.
BspDI  (2146)
1 site
A T C G A T T A G C T A
ClaI  (2146)
1 site
A T C G A T T A G C T A
SalI  (2154)
1 site
G T C G A C C A G C T G
HincII  (2156)
1 site
G T Y R A C C A R Y T G
AbsI  (2160)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2160)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2160)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2160)
1 site
C T C G A G G A G C T C
PspOMI  (2169)
1 site
G G G C C C C C C G G G
ApaI  (2173)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AmpR
3393 .. 4253  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3393 .. 4184  =  792 bp
   263 amino acids  =  29.0 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3393 .. 4253  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4185 .. 4253  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3393 .. 4253  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
HIS3
503 .. 1162  =  660 bp
219 amino acids  =  23.8 kDa
Product: imidazoleglycerol-phosphate dehydratase,
required for histidine biosynthesis
yeast auxotrophic marker
HIS3
503 .. 1162  =  660 bp
219 amino acids  =  23.8 kDa
Product: imidazoleglycerol-phosphate dehydratase,
required for histidine biosynthesis
yeast auxotrophic marker
ori
2634 .. 3222  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
2634 .. 3222  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
lacZ'
1661 .. 2236  =  576 bp
191 amino acids  =  20.8 kDa
Product: fragment of β-galactosidase
lacZ'
1661 .. 2236  =  576 bp
191 amino acids  =  20.8 kDa
Product: fragment of β-galactosidase
CEN/ARS
4395 .. 4898  =  504 bp
S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence
CEN/ARS
4395 .. 4898  =  504 bp
S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence
HIS3 promoter
315 .. 502  =  188 bp
HIS3 promoter
315 .. 502  =  188 bp
AmpR promoter
4254 .. 4358  =  105 bp
AmpR promoter
4254 .. 4358  =  105 bp
lac promoter
2280 .. 2310  =  31 bp
   Segment 3:  -10  
   2280 .. 2286  =  7 bp
promoter for the E. coli lac operon
lac promoter
2280 .. 2310  =  31 bp
   Segment 2:  
   2287 .. 2304  =  18 bp
promoter for the E. coli lac operon
lac promoter
2280 .. 2310  =  31 bp
   Segment 1:  -35  
   2305 .. 2310  =  6 bp
promoter for the E. coli lac operon
lac promoter
2280 .. 2310  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
2256 .. 2272  =  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
2256 .. 2272  =  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).
f1 ori
1423 .. 1878  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1423 .. 1878  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
MCS
2073 .. 2180  =  108 bp
pBluescript multiple cloning site
MCS
2073 .. 2180  =  108 bp
pBluescript multiple cloning site
T7 promoter
2046 .. 2064  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2046 .. 2064  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
2193 .. 2211  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
2193 .. 2211  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
2023 .. 2039  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
2023 .. 2039  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2232 .. 2248  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2232 .. 2248  =  17 bp
common sequencing primer, one of multiple similar
variants
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