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Plasmid Files

pCI

Mammalian cell expression vector with the CMV promoter.

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pCI Sequence and MappCI.dna
Map and Sequence File   
Sequence Author:  Promega
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 BglII (4001) PspFI (3689) BseYI (3685) AlwNI (3580) AhdI (3101) BpmI (3032) NmeAIII (2954) XmnI (2501) EcoO109I (2121) NspI (2076) BsrGI (96) SpeI (152) SnaBI (493) BtgI - NcoI - StyI (513) Eco53kI (719) SacI (721) HindIII (748) PstI (830) BfuAI - BspMI (844) BbsI (928) NheI (1052) BmtI (1056) PaeR7I - XhoI (1058) EcoRI (1063) AflIII - MluI (1069) Acc65I (1075) KpnI (1079) XbaI (1081) SalI (1087) AccI (1088) TspMI - XmaI (1092) SmaI (1094) EagI - NotI (1098) HpaI (1241) MfeI (1250) BspDI - ClaI (1336) BsaBI * (1342) BamHI (1343) NgoMIV (1547) NaeI (1549) DraIII (1655) PfoI (2062) pCI 4006 bp
BglII  (4001)
1 site
A G A T C T T C T A G A
PspFI  (3689)
1 site
C C C A G C G G G T C G
BseYI  (3685)
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.
AlwNI  (3580)
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  (3101)
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  (3032)
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  (2954)
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).
XmnI  (2501)
1 site
G A A N N N N T T C C T T N N N N A A G
EcoO109I  (2121)
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.
NspI  (2076)
1 site
R C A T G Y Y G T A C R
BsrGI  (96)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (152)
1 site
A C T A G T T G A T C A
SnaBI  (493)
1 site
T A C G T A A T G C A T
BtgI  (513)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (513)
1 site
C C A T G G G G T A C C
StyI  (513)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
Eco53kI  (719)
1 site
G A G C T C C T C G A G
SacI  (721)
1 site
G A G C T C C T C G A G
HindIII  (748)
1 site
A A G C T T T T C G A A
PstI  (830)
1 site
C T G C A G G A C G T C
BfuAI  (844)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (844)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BbsI  (928)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
NheI  (1052)
1 site
G C T A G C C G A T C G
BmtI  (1056)
1 site
G C T A G C C G A T C G
PaeR7I  (1058)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1058)
1 site
C T C G A G G A G C T C
EcoRI  (1063)
1 site
G A A T T C C T T A A G
AflIII  (1069)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (1069)
1 site
A C G C G T T G C G C A
Acc65I  (1075)
1 site
G G T A C C C C A T G G
KpnI  (1079)
1 site
G G T A C C C C A T G G
XbaI  (1081)
1 site
T C T A G A A G A T C T
SalI  (1087)
1 site
G T C G A C C A G C T G
AccI  (1088)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
TspMI  (1092)
1 site
C C C G G G G G G C C C
XmaI  (1092)
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  (1094)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EagI  (1098)
1 site
C G G C C G G C C G G C
NotI  (1098)
1 site
G C G G C C G C C G C C G G C G
HpaI  (1241)
1 site
G T T A A C C A A T T G
MfeI  (1250)
1 site
C A A T T G G T T A A C
BspDI  (1336)
1 site
A T C G A T T A G C T A
ClaI  (1336)
1 site
A T C G A T T A G C T A
BsaBI  (1342)
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.
BamHI  (1343)
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.
NgoMIV  (1547)
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  (1549)
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  (1655)
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.
PfoI  (2062)
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.
AmpR
2314 .. 3174  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2314 .. 2382  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2314 .. 3174  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2383 .. 3174  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2314 .. 3174  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3345 .. 3933  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3345 .. 3933  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1422 .. 1877  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1422 .. 1877  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
213 .. 517  =  305 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
213 .. 517  =  305 bp
human cytomegalovirus immediate early enhancer
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
1120 .. 1241  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1120 .. 1241  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2209 .. 2313  =  105 bp
AmpR promoter
2209 .. 2313  =  105 bp
MCS
1052 .. 1104  =  53 bp
multiple cloning site
MCS
1052 .. 1104  =  53 bp
multiple cloning site
T7 promoter
1034 .. 1052  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1034 .. 1052  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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