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Plasmid Files

pCMV-LacZ

Plasmid for high-level expression of E. coli β-galactosidase in mammalian cells. Also known as pCMVβ, pCMVbeta, or pCMV-beta.

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pCMV-LacZ.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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PluTI (7008) SfoI (7006) NarI (7005) KasI (7004) BstAPI (6954) XmnI (6381) ScaI (6262) BsaI (5843) AhdI (5782) BspQI - SapI (4773) HindIII (4530) SalI (4512) MfeI (4446) PsiI (4417) BlpI (3969) BsiWI (3727) EcoRI (1) BsaAI - SnaBI (306) NcoI - StyI (326) BspEI (617) PaeR7I - PspXI - XhoI (640) PpuMI (702) BseRI (730) StuI (769) TspMI - XmaI (830) SmaI (832) Bsu36I (1180) Bpu10I (1461) BspDI - ClaI (1780) EcoRV (2069) DraIII (2146) BsaBI * (2287) BclI * (2302) BssHII - MauBI (2454) AfeI (2791) BstZ17I (3721) pCMV-LacZ 7164 bp
PluTI  (7008)
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.
SfoI  (7006)
1 site
G G C G C C C C G C G G
NarI  (7005)
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.
KasI  (7004)
1 site
G G C G C C C C G C G G
BstAPI  (6954)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
XmnI  (6381)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (6262)
1 site
A G T A C T T C A T G A
BsaI  (5843)
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.
AhdI  (5782)
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.
BspQI  (4773)
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  (4773)
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.
HindIII  (4530)
1 site
A A G C T T T T C G A A
SalI  (4512)
1 site
G T C G A C C A G C T G
MfeI  (4446)
1 site
C A A T T G G T T A A C
PsiI  (4417)
1 site
T T A T A A A A T A T T
BlpI  (3969)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BsiWI  (3727)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
EcoRI  (1)
1 site
G A A T T C C T T A A G
BsaAI  (306)
1 site
Y A C G T R R T G C A Y
SnaBI  (306)
1 site
T A C G T A A T G C A T
NcoI  (326)
1 site
C C A T G G G G T A C C
StyI  (326)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BspEI  (617)
1 site
T C C G G A A G G C C T
PaeR7I  (640)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (640)
1 site
V C T C G A G B B G A G C T C V
XhoI  (640)
1 site
C T C G A G G A G C T C
PpuMI  (702)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BseRI  (730)
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  (769)
1 site
A G G C C T T C C G G A
TspMI  (830)
1 site
C C C G G G G G G C C C
XmaI  (830)
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  (832)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Bsu36I  (1180)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
Bpu10I  (1461)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BspDI  (1780)
1 site
A T C G A T T A G C T A
ClaI  (1780)
1 site
A T C G A T T A G C T A
EcoRV  (2069)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
DraIII  (2146)
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.
BsaBI  (2287)
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.
BclI  (2302)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BssHII  (2454)
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.
MauBI  (2454)
1 site
C G C G C G C G G C G C G C G C
AfeI  (2791)
1 site
A G C G C T T C G C G A
BstZ17I  (3721)
1 site
G T A T A C C A T A T G
lacZ
969 .. 4016  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
969 .. 4016  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
AmpR
5709 .. 6569  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5709 .. 6500  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5709 .. 6569  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6501 .. 6569  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5709 .. 6569  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
4950 .. 5538  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4950 .. 5538  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
4303 .. 4437  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4303 .. 4437  =  135 bp
SV40 polyadenylation signal
AmpR promoter
6570 .. 6674  =  105 bp
AmpR promoter
6570 .. 6674  =  105 bp
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor sites
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor sites
Drosophila Adh
873 .. 965  =  93 bp
31 amino acids  =  3.3 kDa
Product: N-terminal fragment of Drosophila melanogaster alcohol dehydrogenase
Drosophila Adh
873 .. 965  =  93 bp
31 amino acids  =  3.3 kDa
Product: N-terminal fragment of Drosophila melanogaster alcohol dehydrogenase
ORF:  5839 .. 6105  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6955 .. 36  =  246 bp
ORF:  81 amino acids  =  9.5 kDa
ORF:  3890 .. 4174  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  873 .. 4016  =  3144 bp
ORF:  1047 amino acids  =  118.9 kDa
ORF:  4068 .. 4328  =  261 bp
ORF:  86 amino acids  =  10.3 kDa
ORF:  6915 .. 113  =  363 bp
ORF:  120 amino acids  =  13.4 kDa
ORF:  1815 .. 2129  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  3240 .. 3476  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  5709 .. 6569  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2879 .. 3190  =  312 bp
ORF:  103 amino acids  =  10.4 kDa
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