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Plasmid Files

pTEF1/Zeo

Plasmid encoding a yeast expression cassette for a Zeocin™ resistance marker.

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pTEF1 Zeo Sequence and MappTEF1 Zeo.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 PluTI (3319) SfoI (3317) NarI (3316) KasI (3315) PvuII (3246) HindIII (3151) SphI (3149) BfuAI - BspMI (3146) PstI - SbfI (3143) AccI (3134) SalI (3133) XbaI (3127) BamHI (3121) EcoRI (3111) MluI (3030) BsrGI (3023) StuI * (2822) KflI (2814) BseRI (2793) DraIII (2771) FseI (2694) SexAI * (2581) SgrAI (2532) BssHII - MauBI (2454) MscI (2421) NcoI - StyI (2416) BbsI (2239) BtgZI (1991) PaeR7I - PspXI - XhoI (1932) BmtI (1931) NheI (1927) NotI (1914) SacI (1911) Eco53kI (1909) BglII (1901) SfiI (1895) ApaI (1891) PspOMI (1887) AleI (1879) SspI (183) XmnI (388) ScaI (507) TsoI (590) BpmI (919) AhdI (988) AlwNI (1467) PciI (1876) pTEF1/Zeo 3556 bp
PluTI  (3319)
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  (3317)
1 site
G G C G C C C C G C G G
NarI  (3316)
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  (3315)
1 site
G G C G C C C C G C G G
PvuII  (3246)
1 site
C A G C T G G T C G A C
HindIII  (3151)
1 site
A A G C T T T T C G A A
SphI  (3149)
1 site
G C A T G C C G T A C G
BfuAI  (3146)
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  (3146)
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.
PstI  (3143)
1 site
C T G C A G G A C G T C
SbfI  (3143)
1 site
C C T G C A G G G G A C G T C C
AccI  (3134)
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.
SalI  (3133)
1 site
G T C G A C C A G C T G
XbaI  (3127)
1 site
T C T A G A A G A T C T
BamHI  (3121)
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.
EcoRI  (3111)
1 site
G A A T T C C T T A A G
MluI  (3030)
1 site
A C G C G T T G C G C A
BsrGI  (3023)
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.
StuI  (2822)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
KflI  (2814)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
BseRI  (2793)
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.
DraIII  (2771)
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.
FseI  (2694)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
SexAI  (2581)
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.
SgrAI  (2532)
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.
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
MscI  (2421)
1 site
T G G C C A A C C G G T
NcoI  (2416)
1 site
C C A T G G G G T A C C
StyI  (2416)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BbsI  (2239)
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.
BtgZI  (1991)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
PaeR7I  (1932)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1932)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1932)
1 site
C T C G A G G A G C T C
BmtI  (1931)
1 site
G C T A G C C G A T C G
NheI  (1927)
1 site
G C T A G C C G A T C G
NotI  (1914)
1 site
G C G G C C G C C G C C G G C G
SacI  (1911)
1 site
G A G C T C C T C G A G
Eco53kI  (1909)
1 site
G A G C T C C T C G A G
BglII  (1901)
1 site
A G A T C T T C T A G A
SfiI  (1895)
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.
ApaI  (1891)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (1887)
1 site
G G G C C C C C C G G G
AleI  (1879)
1 site
C A C N N N N G T G G T G N N N N C A C
SspI  (183)
1 site
A A T A T T T T A T A A
XmnI  (388)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (507)
1 site
A G T A C T T C A T G A
TsoI  (590)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
BpmI  (919)
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.
AhdI  (988)
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.
AlwNI  (1467)
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.
PciI  (1876)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
201 .. 1061  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   201 .. 269  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
201 .. 1061  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   270 .. 1061  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
201 .. 1061  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1232 .. 1820  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1232 .. 1820  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
TEF1 promoter
1958 .. 2344  =  387 bp
promoter for EF-1α
TEF1 promoter
1958 .. 2344  =  387 bp
promoter for EF-1α
BleoR
2418 .. 2792  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
BleoR
2418 .. 2792  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
CYC1 terminator
2858 .. 3105  =  248 bp
transcription terminator for CYC1
CYC1 terminator
2858 .. 3105  =  248 bp
transcription terminator for CYC1
AmpR promoter
96 .. 200  =  105 bp
AmpR promoter
96 .. 200  =  105 bp
MCS 1
1875 .. 1937  =  63 bp
multiple cloning site 1
MCS 1
1875 .. 1937  =  63 bp
multiple cloning site 1
EM7 promoter
2352 .. 2399  =  48 bp
synthetic bacterial promoter
EM7 promoter
2352 .. 2399  =  48 bp
synthetic bacterial promoter
MCS 2
3111 .. 3156  =  46 bp
multiple cloning site 2
MCS 2
3111 .. 3156  =  46 bp
multiple cloning site 2
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