Resources
Plasmid Files

pRS423

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

 
To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

 MfeI (5679) XcmI (5678) BmgBI (5467) SnaBI (5029) BsaHI (4006) ScaI (3949) NmeAIII (3617) BsaI (3530) AhdI (3469) AlwNI (2992) PspFI (2884) PfoI (46) BsaBI * (561) NdeI (694) MscI (711) BsmI (716) BsiWI (916) NheI (1008) BmtI (1012) BclI * (1052) PsiI (1519) DraIII (1647) BtgZI (1648) NgoMIV (1748) NaeI (1750) PspOMI (2082) ApaI (2086) AbsI - PaeR7I - PspXI - XhoI (2091) SalI (2097) HincII (2099) BspDI - ClaI (2107) EcoRV (2120) EcoRI (2124) TspMI - XmaI (2136) SmaI (2138) BamHI (2142) SpeI (2148) EagI - NotI (2161) BtgI (2170) SacII (2173) Eco53kI (2180) SacI (2182) lac operator BspQI - SapI (2460) BseYI (2880) pRS423 5797 bp
MfeI  (5679)
1 site
C A A T T G G T T A A C
XcmI  (5678)
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.
BmgBI  (5467)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
SnaBI  (5029)
1 site
T A C G T A A T G C A T
BsaHI  (4006)
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  (3949)
1 site
A G T A C T T C A T G A
NmeAIII  (3617)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaI  (3530)
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  (3469)
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  (2992)
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  (2884)
1 site
C C C A G C G G G T C 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.
BsaBI  (561)
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.
NdeI  (694)
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.
MscI  (711)
1 site
T G G C C A A C C G G T
BsmI  (716)
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  (916)
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  (1008)
1 site
G C T A G C C G A T C G
BmtI  (1012)
1 site
G C T A G C C G A T C G
BclI  (1052)
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.
PsiI  (1519)
1 site
T T A T A A A A T A T T
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.
PspOMI  (2082)
1 site
G G G C C C C C C G G G
ApaI  (2086)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AbsI  (2091)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2091)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2091)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2091)
1 site
C T C G A G G A G C T C
SalI  (2097)
1 site
G T C G A C C A G C T G
HincII  (2099)
1 site
G T Y R A C C A R Y T G
BspDI  (2107)
1 site
A T C G A T T A G C T A
ClaI  (2107)
1 site
A T C G A T T A G C T A
EcoRV  (2120)
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.
EcoRI  (2124)
1 site
G A A T T C C T T A A G
TspMI  (2136)
1 site
C C C G G G G G G C C C
XmaI  (2136)
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  (2138)
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  (2142)
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  (2148)
1 site
A C T A G T T G A T C A
EagI  (2161)
1 site
C G G C C G G C C G G C
NotI  (2161)
1 site
G C G G C C G C C G C C G G C G
BtgI  (2170)
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  (2173)
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.
Eco53kI  (2180)
1 site
G A G C T C C T C G A G
SacI  (2182)
1 site
G A G C T C C T C G A G
BspQI  (2460)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2460)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
BseYI  (2880)
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.
2μ ori
4388 .. 5730  =  1343 bp
yeast 2μ plasmid origin of replication
2μ ori
4388 .. 5730  =  1343 bp
yeast 2μ plasmid origin of replication
AmpR
3396 .. 4256  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3396 .. 4187  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3396 .. 4256  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4188 .. 4256  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3396 .. 4256  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
HIS3
504 .. 1163  =  660 bp
219 amino acids  =  23.8 kDa
Product: imidazoleglycerol-phosphate dehydratase,
required for histidine biosynthesis
yeast auxotrophic marker
HIS3
504 .. 1163  =  660 bp
219 amino acids  =  23.8 kDa
Product: imidazoleglycerol-phosphate dehydratase,
required for histidine biosynthesis
yeast auxotrophic marker
ori
2637 .. 3225  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
2637 .. 3225  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
lacZα
1661 .. 2239  =  579 bp
192 amino acids  =  20.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
1661 .. 2239  =  579 bp
192 amino acids  =  20.6 kDa
Product: LacZα fragment of β-galactosidase
HIS3 promoter
317 .. 503  =  187 bp
HIS3 promoter
317 .. 503  =  187 bp
AmpR promoter
4257 .. 4361  =  105 bp
AmpR promoter
4257 .. 4361  =  105 bp
lac promoter
2283 .. 2313  =  31 bp
   Segment 3:  -10  
   2283 .. 2289  =  7 bp
promoter for the E. coli lac operon
lac promoter
2283 .. 2313  =  31 bp
   Segment 2:  
   2290 .. 2307  =  18 bp
promoter for the E. coli lac operon
lac promoter
2283 .. 2313  =  31 bp
   Segment 1:  -35  
   2308 .. 2313  =  6 bp
promoter for the E. coli lac operon
lac promoter
2283 .. 2313  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
2259 .. 2275  =  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
2259 .. 2275  =  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
2076 .. 2183  =  108 bp
pBluescript multiple cloning site
MCS
2076 .. 2183  =  108 bp
pBluescript multiple cloning site
T7 promoter
2049 .. 2067  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2049 .. 2067  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
2196 .. 2214  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
2196 .. 2214  =  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
2235 .. 2251  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2235 .. 2251  =  17 bp
common sequencing primer, one of multiple similar
variants
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter