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Plasmid Files

pRS415

Yeast centromere vector with a LEU2 marker and an MCS derived from pBLUESCRIPT II.

 
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 SwaI (5477) PmlI (5453) ScaI (5000) BsaI (4581) AhdI (4520) AlwNI (4043) PspFI (3935) BseYI (3931) lac operator SacI (3233) Eco53kI (3231) SacII (3224) AleI (3223) EagI - NotI (3212) XbaI (3205) SpeI (3199) BamHI (3193) SmaI (3189) TspMI - XmaI (3187) PstI (3185) HindIII (3163) SalI (3148) AbsI - PaeR7I - PspXI - XhoI (3142) ApaI (3137) PspOMI (3133) PfoI (46) PflFI - Tth111I (192) BsrGI (632) BfuAI - BspMI (759) XcmI (1201) AflII (1438) AgeI (1503) BstEII (1718) HpaI (2174) KasI (2203) NarI (2204) SfoI (2205) PluTI (2207) DraIII (2698) BtgZI (2699) NgoMIV (2799) NaeI (2801) pRS415 6021 bp
SwaI  (5477)
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  (5453)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
ScaI  (5000)
1 site
A G T A C T T C A T G A
BsaI  (4581)
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  (4520)
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  (4043)
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.
PspFI  (3935)
1 site
C C C A G C G G G T C G
BseYI  (3931)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
SacI  (3233)
1 site
G A G C T C C T C G A G
Eco53kI  (3231)
1 site
G A G C T C C T C G A G
SacII  (3224)
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.
AleI  (3223)
1 site
C A C N N N N G T G G T G N N N N C A C
EagI  (3212)
1 site
C G G C C G G C C G G C
NotI  (3212)
1 site
G C G G C C G C C G C C G G C G
XbaI  (3205)
1 site
T C T A G A A G A T C T
SpeI  (3199)
1 site
A C T A G T T G A T C A
BamHI  (3193)
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  (3189)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (3187)
1 site
C C C G G G G G G C C C
XmaI  (3187)
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.
PstI  (3185)
1 site
C T G C A G G A C G T C
HindIII  (3163)
1 site
A A G C T T T T C G A A
SalI  (3148)
1 site
G T C G A C C A G C T G
AbsI  (3142)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (3142)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3142)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3142)
1 site
C T C G A G G A G C T C
ApaI  (3137)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (3133)
1 site
G G G C C C C C C G G G
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.
PflFI  (192)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (192)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsrGI  (632)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
BfuAI  (759)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (759)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
XcmI  (1201)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
AflII  (1438)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
AgeI  (1503)
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.
BstEII  (1718)
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.
HpaI  (2174)
1 site
G T T A A C C A A T T G
KasI  (2203)
1 site
G G C G C C C C G C G G
NarI  (2204)
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  (2205)
1 site
G G C G C C C C G C G G
PluTI  (2207)
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.
DraIII  (2698)
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  (2699)
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  (2799)
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  (2801)
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.
LEU2
663 .. 1757  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required
for leucine biosynthesis
yeast auxotrophic marker
LEU2
663 .. 1757  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required
for leucine biosynthesis
yeast auxotrophic marker
AmpR
4447 .. 5307  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4447 .. 5238  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4447 .. 5307  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5239 .. 5307  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4447 .. 5307  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3688 .. 4276  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
3688 .. 4276  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
lacZα
2712 .. 3290  =  579 bp
192 amino acids  =  20.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2712 .. 3290  =  579 bp
192 amino acids  =  20.6 kDa
Product: LacZα fragment of β-galactosidase
CEN/ARS
5449 .. 5952  =  504 bp
S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence
CEN/ARS
5449 .. 5952  =  504 bp
S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence
LEU2 promoter
1770 .. 2174  =  405 bp
LEU2 promoter
1770 .. 2174  =  405 bp
AmpR promoter
5308 .. 5412  =  105 bp
AmpR promoter
5308 .. 5412  =  105 bp
lac promoter
3334 .. 3364  =  31 bp
   Segment 3:  -10  
   3334 .. 3340  =  7 bp
promoter for the E. coli lac operon
lac promoter
3334 .. 3364  =  31 bp
   Segment 2:  
   3341 .. 3358  =  18 bp
promoter for the E. coli lac operon
lac promoter
3334 .. 3364  =  31 bp
   Segment 1:  -35  
   3359 .. 3364  =  6 bp
promoter for the E. coli lac operon
lac promoter
3334 .. 3364  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
3310 .. 3326  =  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
3310 .. 3326  =  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
2474 .. 2929  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2474 .. 2929  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
MCS
3127 .. 3234  =  108 bp
pBluescript multiple cloning site
MCS
3127 .. 3234  =  108 bp
pBluescript multiple cloning site
T7 promoter
3100 .. 3118  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3100 .. 3118  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
3247 .. 3265  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
3247 .. 3265  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
3074 .. 3090  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
3074 .. 3090  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
3286 .. 3302  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
3286 .. 3302  =  17 bp
common sequencing primer, one of multiple similar
variants
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