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Plasmid Files

pCEP4

Episomal mammalian vector with a CMV promoter and a hygromycin resistance marker, for high-level expression in primate and canine cells.

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pCEP4.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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BsrGI (10,148) AsiSI (8773) PshAI (8436) BstBI (8260) NruI (7959) XmnI (6358) BstEII (5867) EcoNI (5862) StuI * (5791) AvrII (5502) SnaBI (359) Acc65I (620) KpnI (624) PvuII (627) HindIII (636) NotI (647) PaeR7I - PspXI - XhoI (653) NgoMIV (659) NaeI (661) SfiI (665) BamHI (671) EcoRV (1661) NsiI (3212) SexAI * (3386) BbvCI (3827) SgrAI (4028) pCEP4 10,186 bp
BsrGI  (10,148)
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.
AsiSI  (8773)
1 site
G C G A T C G C C G C T A G C G
PshAI  (8436)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BstBI  (8260)
1 site
T T C G A A A A G C T T
NruI  (7959)
1 site
T C G C G A A G C G C T
XmnI  (6358)
1 site
G A A N N N N T T C C T T N N N N A A G
BstEII  (5867)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
EcoNI  (5862)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
StuI  (5791)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
AvrII  (5502)
1 site
C C T A G G G G A T C C
SnaBI  (359)
1 site
T A C G T A A T G C A T
Acc65I  (620)
1 site
G G T A C C C C A T G G
KpnI  (624)
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
HindIII  (636)
1 site
A A G C T T T T C G A A
NotI  (647)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (653)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (653)
1 site
V C T C G A G B B G A G C T C V
XhoI  (653)
1 site
C T C G A G G A G C T C
NgoMIV  (659)
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  (661)
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.
SfiI  (665)
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.
BamHI  (671)
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.
EcoRV  (1661)
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.
NsiI  (3212)
1 site
A T G C A T T A C G T A
SexAI  (3386)
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.
BbvCI  (3827)
1 site
C C T C A G C G G A G T C G
SgrAI  (4028)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
EBNA1
3620 .. 5545  =  1926 bp
641 amino acids  =  56.4 kDa
Product: Epstein-Barr nuclear antigen 1, also known as EBNA-1
crucial for latent viral infection, and for episomal amplification of vectors containing the oriP origin (Okita et al., 2013)
EBNA1
3620 .. 5545  =  1926 bp
641 amino acids  =  56.4 kDa
Product: Epstein-Barr nuclear antigen 1, also known as EBNA-1
crucial for latent viral infection, and for episomal amplification of vectors containing the oriP origin (Okita et al., 2013)
oriP
1529 .. 3318  =  1790 bp
Epstein-Barr virus oriP replication origin (Yates et al., 2000)
oriP
1529 .. 3318  =  1790 bp
Epstein-Barr virus oriP replication origin (Yates et al., 2000)
HygR
8409 .. 9446  =  1038 bp
345 amino acids  =  38.5 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
8409 .. 9446  =  1038 bp
345 amino acids  =  38.5 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
AmpR
6171 .. 7031  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   6171 .. 6239  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6171 .. 7031  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   6240 .. 7031  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6171 .. 7031  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7202 .. 7790  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7202 .. 7790  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
4 .. 383  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4 .. 383  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
384 .. 587  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
384 .. 587  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
HSV TK promoter
8221 .. 8366  =  146 bp
herpes simplex virus thymidine kinase promoter
HSV TK promoter
8221 .. 8366  =  146 bp
herpes simplex virus thymidine kinase promoter
SV40 poly(A) signal
672 .. 806  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
672 .. 806  =  135 bp
SV40 polyadenylation signal
AmpR promoter
6066 .. 6170  =  105 bp
AmpR promoter
6066 .. 6170  =  105 bp
HSV TK poly(A) signal
9488 .. 9536  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
9488 .. 9536  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
MCS
620 .. 676  =  57 bp
multiple cloning site
MCS
620 .. 676  =  57 bp
multiple cloning site
ORF:  4333 .. 5547  =  1215 bp
ORF:  404 amino acids  =  45.2 kDa
ORF:  9736 .. 10,029  =  294 bp
ORF:  97 amino acids  =  11.1 kDa
ORF:  1061 .. 1471  =  411 bp
ORF:  136 amino acids  =  14.7 kDa
ORF:  4142 .. 4378  =  237 bp
ORF:  78 amino acids  =  8.9 kDa
ORF:  7760 .. 8092  =  333 bp
ORF:  110 amino acids  =  12.3 kDa
ORF:  9743 .. 10,051  =  309 bp
ORF:  102 amino acids  =  11.7 kDa
ORF:  6171 .. 7031  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  8409 .. 9446  =  1038 bp
ORF:  345 amino acids  =  38.5 kDa
ORF:  3620 .. 5545  =  1926 bp
ORF:  641 amino acids  =  56.4 kDa
ORF:  6635 .. 6901  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7709 .. 7945  =  237 bp
ORF:  78 amino acids  =  8.9 kDa
ORF:  1021 .. 1254  =  234 bp
ORF:  77 amino acids  =  8.3 kDa
ORF:  3220 .. 3537  =  318 bp
ORF:  105 amino acids  =  11.7 kDa
ORF:  4318 .. 5430  =  1113 bp
ORF:  370 amino acids  =  40.7 kDa
ORF:  7465 .. 7863  =  399 bp
ORF:  132 amino acids  =  14.8 kDa
ORF:  7927 .. 8205  =  279 bp
ORF:  92 amino acids  =  10.5 kDa
ORF:  2241 .. 2480  =  240 bp
ORF:  79 amino acids  =  8.7 kDa
ORF:  8070 .. 9737  =  1668 bp
ORF:  555 amino acids  =  60.8 kDa
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