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Plasmid Files

pRS315

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

 
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pRS315.dna
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SwaI (5474) PmlI (5450) ScaI (4997) BsaI (4578) AhdI (4517) AlwNI (4040) ApaI (3224) PspOMI (3220) SalI (3205) HindIII (3190) PstI (3176) SmaI (3168) TspMI - XmaI (3166) BamHI (3160) SpeI (3154) XbaI (3148) EagI - NotI (3141) SacII (3135) AleI (3134) SacI (3128) Eco53kI (3126) 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) pRS315 6018 bp
SwaI  (5474)
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  (5450)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
ScaI  (4997)
1 site
A G T A C T T C A T G A
BsaI  (4578)
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  (4517)
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  (4040)
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.
ApaI  (3224)
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  (3220)
1 site
G G G C C C C C C G G G
SalI  (3205)
1 site
G T C G A C C A G C T G
HindIII  (3190)
1 site
A A G C T T T T C G A A
PstI  (3176)
1 site
C T G C A G G A C G T C
SmaI  (3168)
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  (3166)
1 site
C C C G G G G G G C C C
XmaI  (3166)
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.
BamHI  (3160)
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.
SpeI  (3154)
1 site
A C T A G T T G A T C A
XbaI  (3148)
1 site
T C T A G A A G A T C T
EagI  (3141)
1 site
C G G C C G G C C G G C
NotI  (3141)
1 site
G C G G C C G C C G C C G G C G
SacII  (3135)
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  (3134)
1 site
C A C N N N N G T G G T G N N N N C A C
SacI  (3128)
1 site
G A G C T C C T C G A G
Eco53kI  (3126)
1 site
G A G C T C C T C G A 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
AgeI  (1503)
1 site
A C C G G T T G G C C A
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
4444 .. 5304  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4444 .. 5235  =  792 bp
   263 amino acids  =  29.0 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4444 .. 5304  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5236 .. 5304  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4444 .. 5304  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3685 .. 4273  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
3685 .. 4273  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
lacZ'
2712 .. 3287  =  576 bp
191 amino acids  =  20.8 kDa
Product: fragment of β-galactosidase
lacZ'
2712 .. 3287  =  576 bp
191 amino acids  =  20.8 kDa
Product: fragment of β-galactosidase
CEN/ARS
5446 .. 5949  =  504 bp
S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence
CEN/ARS
5446 .. 5949  =  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
5305 .. 5409  =  105 bp
AmpR promoter
5305 .. 5409  =  105 bp
lac promoter
3331 .. 3361  =  31 bp
   Segment 3:  -10  
   3331 .. 3337  =  7 bp
promoter for the E. coli lac operon
lac promoter
3331 .. 3361  =  31 bp
   Segment 2:  
   3338 .. 3355  =  18 bp
promoter for the E. coli lac operon
lac promoter
3331 .. 3361  =  31 bp
   Segment 1:  -35  
   3356 .. 3361  =  6 bp
promoter for the E. coli lac operon
lac promoter
3331 .. 3361  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
3307 .. 3323  =  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
3307 .. 3323  =  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
3124 .. 3231  =  108 bp
pBluescript multiple cloning site
MCS
3124 .. 3231  =  108 bp
pBluescript multiple cloning site
T7 promoter
3097 .. 3115  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3097 .. 3115  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
3244 .. 3262  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
3244 .. 3262  =  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
3283 .. 3299  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
3283 .. 3299  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  784 .. 1095  =  312 bp
ORF:  103 amino acids  =  11.1 kDa
ORF:  4574 .. 4840  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4444 .. 5304  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  663 .. 1769  =  1107 bp
ORF:  368 amino acids  =  39.4 kDa
ORF:  2712 .. 3287  =  576 bp
ORF:  191 amino acids  =  20.8 kDa
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