pRKW2

Mammalian expression vector for cDNA cloning.

Sequence Author: I.M.A.G.E. Consortium

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HomePlasmidsI.M.A.G.E. Consortium PlasmidspRKW2
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) CsiI - 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
CsiI  (1324)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
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
2 segments
   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
2 segments
   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
ORF:  553 .. 798  =  246 bp
ORF:  81 amino acids  =  9.8 kDa
ORF:  2713 .. 3573  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1484 .. 1801  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  3177 .. 3443  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Individual Sequences & Maps

Lafmid BA
pcDNAII
pCR2.1-TOPO (linearized)
pExpress-1
pAD-GAL4
pcDNA3.1(+)
pCR3.1
pME18S-FL3
pAMP1
pcDNA3.1(-)
pCR3.1 (linearized)
pOTB7
pAMP10
pcDNA3.1 Zeo(+)
pCR4-TOPO
pOTB7-3
pBK-CMV
pCMV SPORT
pCR4-TOPO (linearized)
pOTB7a
pBluescribe (modified)
pCMV SPORT2
pCS105
pPCR-Script Amp SK(+)
pBluescript II SK(+)
pCMV SPORT4
pCS107
pRKW2
pBluescript KS(+)
pCMV SPORT6
pCS108
pSPORT 1
pBluescript SK(+)
pCMV SPORT6.1
pCS2+
pSPORT 2
pBluescript SK(-)
pCMV SPORT6ccdB
pCS22+
pT7T3D-PacI
pBluescript-FL
pCR-Blunt II-TOPO
pCS2G
pUC19
pBluescript II KS(+)
pCR-Blunt II-TOPO (linearized)
pDNR-Dual
pYX-Asc
pBluescriptR
pCR-XL-TOPO
pDNR-LIB
pZErO-2
pcDNA3
pCR-XL-TOPO (linearized)
pDONR201
pZL1
pcDNAI
pCR2.1-TOPO
pDONR223

Plasmid Sets

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TA and GC Cloning Vectors
Coronavirus Resources
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TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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