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Plasmid Files

pCS105

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

 
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 HindIII (28) NdeI (3697) MscI * (3510) BseRI (3389) NaeI (2941) NgoMIV (2939) DraIII (2838) XmnI (2295) SP6 promoter BamHI (79) BspDI - ClaI (87) BstBI (92) EcoRI (94) SalI (100) SfiI (114) EagI - NotI (121) StuI (131) AvaI - BsoBI - PaeR7I - XhoI (135) BmeT110I (136) XbaI (143) SnaBI (168) HpaI (306) MfeI (315) AscI (376) BstXI - NsiI (396) PspOMI (399) ApaI (403) Acc65I (405) KpnI (409) M13 rev lac operator PvuII (627) BspQI - SapI (687) AflIII - PciI (803) NspI (807) AlwNI (1219) AhdI (1696) BpmI (1766) NmeAIII (1844) pCS105 4131 bp
HindIII  (28)
1 site
A A G C T T T T C G A A
NdeI  (3697)
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  (3510)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BseRI  (3389)
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.
NaeI  (2941)
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  (2939)
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  (2838)
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  (2295)
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
SalI  (100)
1 site
G T C G A C C A G C T G
SfiI  (114)
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.
EagI  (121)
1 site
C G G C C G G C C G G C
NotI  (121)
1 site
G C G G C C G C C G C C G G C G
StuI  (131)
1 site
A G G C C T T C C G G A
AvaI  (135)
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  (135)
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  (135)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (135)
1 site
C T C G A G G A G C T C
BmeT110I  (136)
1 site
C Y C G R G G R G C Y C
XbaI  (143)
1 site
T C T A G A A G A T C T
SnaBI  (168)
1 site
T A C G T A A T G C A T
HpaI  (306)
1 site
G T T A A C C A A T T G
MfeI  (315)
1 site
C A A T T G G T T A A C
AscI  (376)
1 site
G G C G C G C C C C G C G C G G
BstXI  (396)
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  (396)
1 site
A T G C A T T A C G T A
PspOMI  (399)
1 site
G G G C C C C C C G G G
ApaI  (403)
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  (405)
1 site
G G T A C C C C A T G G
KpnI  (409)
1 site
G G T A C C C C A T G G
PvuII  (627)
1 site
C A G C T G G T C G A C
BspQI  (687)
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  (687)
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  (803)
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  (803)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (807)
1 site
R C A T G Y Y G T A C R
AlwNI  (1219)
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  (1696)
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  (1766)
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  (1844)
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
3147 .. 4131  =  985 bp
enhancer/promoter region of simian
cytomegalovirus major immediate early
transcription unit IE94
CMV IE94 promoter
3147 .. 4131  =  985 bp
enhancer/promoter region of simian
cytomegalovirus major immediate early
transcription unit IE94
AmpR
1623 .. 2483  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1623 .. 2414  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1623 .. 2483  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2415 .. 2483  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1623 .. 2483  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
864 .. 1452  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
864 .. 1452  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
2614 .. 3069  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2614 .. 3069  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 poly(A) signal
172 .. 306  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
172 .. 306  =  135 bp
SV40 polyadenylation signal
AmpR promoter
2484 .. 2588  =  105 bp
AmpR promoter
2484 .. 2588  =  105 bp
lac promoter
510 .. 540  =  31 bp
   Segment 3:  -10  
   510 .. 516  =  7 bp
promoter for the E. coli lac operon
lac promoter
510 .. 540  =  31 bp
   Segment 2:  
   517 .. 534  =  18 bp
promoter for the E. coli lac operon
lac promoter
510 .. 540  =  31 bp
   Segment 1:  -35  
   535 .. 540  =  6 bp
promoter for the E. coli lac operon
lac promoter
510 .. 540  =  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
423 .. 441  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
423 .. 441  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 rev
462 .. 478  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
462 .. 478  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
486 .. 502  =  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
486 .. 502  =  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
145 .. 172  =  28 bp
stop codons in all three reading frames
stop codons
145 .. 172  =  28 bp
stop codons in all three reading frames
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