Resources
Plasmid Files

pRKW2

Mammalian expression vector for cDNA cloning.

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

pRKW2 Sequence and MappRKW2.dna
Map and Sequence File   
Sequence Author:  I.M.A.G.E. Consortium
Download Free Trial Get SnapGene Viewer

 BspQI - SapI (4505) AflIII - PciI (4388) AlwNI (3979) AhdI (3500) BsaI (3434) FspI (3277) ScaI (3019) EcoO109I (2520) PfoI (2446) NdeI (263) SnaBI (369) AG195 (822 .. 845) BsgI (882) PspOMI (916) ApaI (920) PaeR7I - PspXI - XhoI (949) BamHI (955) XbaI (967) NotI (974) AleI (985) AG189 (1051 .. 1074) BsmI (1088) BspDI * - ClaI * (1141) Acc65I (1207) KpnI (1211) SexAI * (1324) SfiI (1510) BseRI (1553) StuI (1556) AvrII (1557) HincII - HpaI (1580) KasI (1747) NarI (1748) SfoI (1749) PluTI (1751) NgoMIV (1946) NaeI (1948) DraIII (2054) PflFI - Tth111I (2347) pRKW2 4750 bp
BspQI  (4505)
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  (4505)
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.
AflIII  (4388)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4388)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (3979)
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.
AhdI  (3500)
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.
BsaI  (3434)
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.
FspI  (3277)
1 site
T G C G C A A C G C G T
ScaI  (3019)
1 site
A G T A C T T C A T G A
EcoO109I  (2520)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PfoI  (2446)
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.
NdeI  (263)
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.
SnaBI  (369)
1 site
T A C G T A A T G C A T
BsgI  (882)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PspOMI  (916)
1 site
G G G C C C C C C G G G
ApaI  (920)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PaeR7I  (949)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (949)
1 site
V C T C G A G B B G A G C T C V
XhoI  (949)
1 site
C T C G A G G A G C T C
BamHI  (955)
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  (967)
1 site
T C T A G A A G A T C T
NotI  (974)
1 site
G C G G C C G C C G C C G G C G
AleI  (985)
1 site
C A C N N N N G T G G T G N N N N C A C
BsmI  (1088)
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.
BspDI  (1141)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1141)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
Acc65I  (1207)
1 site
G G T A C C C C A T G G
KpnI  (1211)
1 site
G G T A C C C C A T G G
SexAI  (1324)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (1510)
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.
BseRI  (1553)
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.
StuI  (1556)
1 site
A G G C C T T C C G G A
AvrII  (1557)
1 site
C C T A G G G G A T C C
HincII  (1580)
1 site
G T Y R A C C A R Y T G
HpaI  (1580)
1 site
G T T A A C C A A T T G
KasI  (1747)
1 site
G G C G C C C C G C G G
NarI  (1748)
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  (1749)
1 site
G G C G C C C C G C G G
PluTI  (1751)
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.
NgoMIV  (1946)
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  (1948)
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.
DraIII  (2054)
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.
PflFI  (2347)
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  (2347)
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.
AG195
24-mer  /  38% GC
1 binding site
822 .. 845  =  24 annealed bases
Tm  =  52°C
AG189
24-mer  /  29% GC
1 binding site
1051 .. 1074  =  24 annealed bases
Tm  =  52°C
AmpR
2713 .. 3573  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   2713 .. 2781  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2713 .. 3573  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2782 .. 3573  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2713 .. 3573  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3744 .. 4332  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3744 .. 4332  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1821 .. 2276  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1821 .. 2276  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
14 .. 393  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
14 .. 393  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
394 .. 597  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
394 .. 597  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 ori
1423 .. 1558  =  136 bp
SV40 origin of replication
SV40 ori
1423 .. 1558  =  136 bp
SV40 origin of replication
SV40 poly(A) signal
1008 .. 1142  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1008 .. 1142  =  135 bp
SV40 polyadenylation signal
AmpR promoter
2608 .. 2712  =  105 bp
AmpR promoter
2608 .. 2712  =  105 bp
SP6 promoter
828 .. 846  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
828 .. 846  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 fwd
1592 .. 1608  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
1592 .. 1608  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
4718 .. 4734  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
4718 .. 4734  =  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