pCS2+

Vector that allows high-level transient expression in vertebrate cells as well as in vitro transcription/translation.

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

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HomePlasmidsI.M.A.G.E. Consortium PlasmidspCS2+
HindIII (28) NdeI (3661) MscI * (3474) BseRI (3353) SalI (3108) NaeI (2909) NgoMIV (2907) DraIII (2806) XmnI (2263) SP6 promoter BamHI (79) BspDI - ClaI (87) BstBI (92) EcoRI (94) StuI (103) AvaI - BsoBI - PaeR7I - XhoI (107) XbaI (115) SnaBI (140) HpaI (278) MfeI (287) EagI - NotI (346) BtgI (349) SacII (352) BstXI - NsiI (364) PspOMI (367) ApaI (371) Acc65I (373) KpnI (377) BssHII (410) M13 rev lac operator PvuII (595) BspQI - SapI (655) AflIII - PciI (771) NspI (775) AlwNI (1187) AhdI (1664) BpmI (1734) NmeAIII (1812) pCS2+ 4095 bp
HindIII  (28)
1 site		 A A G C T T T T C G A A
NdeI  (3661)
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  (3474)
1 site		 T G G C C A A C C G G T
* Blocked by Dcm methylation.
BseRI  (3353)
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.
SalI  (3108)
1 site		 G T C G A C C A G C T G
NaeI  (2909)
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.
NgoMIV  (2907)
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.
DraIII  (2806)
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.
XmnI  (2263)
1 site		 G A A N N N N T T C C T T N N N N A A G
BamHI  (79)
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.
BspDI  (87)
1 site		 A T C G A T T A G C T A
ClaI  (87)
1 site		 A T C G A T T A G C T A
BstBI  (92)
1 site		 T T C G A A A A G C T T
EcoRI  (94)
1 site		 G A A T T C C T T A A G
StuI  (103)
1 site		 A G G C C T T C C G G A
AvaI  (107)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (107)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (107)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (107)
1 site		 C T C G A G G A G C T C
XbaI  (115)
1 site		 T C T A G A A G A T C T
SnaBI  (140)
1 site		 T A C G T A A T G C A T
HpaI  (278)
1 site		 G T T A A C C A A T T G
MfeI  (287)
1 site		 C A A T T G G T T A A C
EagI  (346)
1 site		 C G G C C G G C C G G C
NotI  (346)
1 site		 G C G G C C G C C G C C G G C G
BtgI  (349)
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  (352)
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.
BstXI  (364)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
NsiI  (364)
1 site		 A T G C A T T A C G T A
PspOMI  (367)
1 site		 G G G C C C C C C G G G
ApaI  (371)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
Acc65I  (373)
1 site		 G G T A C C C C A T G G
KpnI  (377)
1 site		 G G T A C C C C A T G G
BssHII  (410)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
PvuII  (595)
1 site		 C A G C T G G T C G A C
BspQI  (655)
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  (655)
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  (771)
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  (771)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
NspI  (775)
1 site		 R C A T G Y Y G T A C R
AlwNI  (1187)
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  (1664)
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.
BpmI  (1734)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
NmeAIII  (1812)
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).
CMV IE94 promoter
3111 .. 4095  =  985 bp
enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94
CMV IE94 promoter
3111 .. 4095  =  985 bp
enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94
AmpR
1591 .. 2451  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1591 .. 2382  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1591 .. 2451  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2383 .. 2451  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1591 .. 2451  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
832 .. 1420  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
832 .. 1420  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2582 .. 3037  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2582 .. 3037  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 poly(A) signal
144 .. 278  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
144 .. 278  =  135 bp
SV40 polyadenylation signal
AmpR promoter
2452 .. 2556  =  105 bp
AmpR promoter
2452 .. 2556  =  105 bp
lac promoter
478 .. 508  =  31 bp
3 segments
   Segment 3:  -10  
   478 .. 484  =  7 bp
promoter for the E. coli lac operon
lac promoter
478 .. 508  =  31 bp
3 segments
   Segment 2:  
   485 .. 502  =  18 bp
promoter for the E. coli lac operon
lac promoter
478 .. 508  =  31 bp
3 segments
   Segment 1:  -35  
   503 .. 508  =  6 bp
promoter for the E. coli lac operon
lac promoter
478 .. 508  =  31 bp
3 segments
promoter for the E. coli lac operon
SP6 promoter
35 .. 53  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
35 .. 53  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
T3 promoter
391 .. 409  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
391 .. 409  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 rev
430 .. 446  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
430 .. 446  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
454 .. 470  =  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
454 .. 470  =  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).
stop codons
117 .. 144  =  28 bp
stop codons in all three reading frames
stop codons
117 .. 144  =  28 bp
stop codons in all three reading frames
ORF:  1721 .. 1987  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3146 .. 3544  =  399 bp
ORF:  132 amino acids  =  14.3 kDa
ORF:  3129 .. 3377  =  249 bp
ORF:  82 amino acids  =  9.1 kDa
ORF:  3777 .. 4049  =  273 bp
ORF:  90 amino acids  =  9.5 kDa
ORF:  1591 .. 2451  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2820 .. 3116  =  297 bp
ORF:  98 amino acids  =  10.6 kDa
ORF:  3050 .. 3406  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  3749 .. 4030  =  282 bp
ORF:  93 amino acids  =  10.1 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
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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