pETcoco-1

Bacterial vector with a chloramphenicol resistance marker that allows single-copy replication, on-demand amplification, and tightly regulated expression.

Sequence Author: MilliporeSigma (Novagen)

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lac operator T7 promoter ApaLI (11,732) PciI (10,208) BsaI (9536) SpeI (9448) BseRI (8426) SnaBI (8357) PflFI - Tth111I (7997) KpnI (7937) Acc65I (7933) SbfI (6751) BsrGI (6506) FspAI (6478) RBS SphI (74) NsiI (76) NheI (96) BmtI (100) SrfI (181) AsiSI (188) ZraI (200) AatII (202) AscI (207) PmlI (215) BsiWI (219) BstBI (228) HindIII (231) NotI (238) PacI (288) Bsu36I (293) SwaI (302) AvrII (309) PspXI (320) 6xHis BamHI (1974) SfiI (2434) PaqCI (2679) ScaI (3424) BstZ17I (4463) FseI (5624) SacII (5628) StuI (5900) pETcoco™-1 12,474 bp
ApaLI  (11,732)
1 site
G T G C A C C A C G T G
PciI  (10,208)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BsaI  (9536)
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.
SpeI  (9448)
1 site
A C T A G T T G A T C A
BseRI  (8426)
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.
SnaBI  (8357)
1 site
T A C G T A A T G C A T
PflFI  (7997)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (7997)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
KpnI  (7937)
1 site
G G T A C C C C A T G G
Acc65I  (7933)
1 site
G G T A C C C C A T G G
SbfI  (6751)
1 site
C C T G C A G G G G A C G T C C
BsrGI  (6506)
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.
FspAI  (6478)
1 site
R T G C G C A Y Y A C G C G T R
SphI  (74)
1 site
G C A T G C C G T A C G
NsiI  (76)
1 site
A T G C A T T A C G T A
NheI  (96)
1 site
G C T A G C C G A T C G
BmtI  (100)
1 site
G C T A G C C G A T C G
SrfI  (181)
1 site
G C C C G G G C C G G G C C C G
AsiSI  (188)
1 site
G C G A T C G C C G C T A G C G
ZraI  (200)
1 site
G A C G T C C T G C A G
AatII  (202)
1 site
G A C G T C C T G C A G
AscI  (207)
1 site
G G C G C G C C C C G C G C G G
PmlI  (215)
1 site
C A C G T G G T G C A C
BsiWI  (219)
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.
BstBI  (228)
1 site
T T C G A A A A G C T T
HindIII  (231)
1 site
A A G C T T T T C G A A
NotI  (238)
1 site
G C G G C C G C C G C C G G C G
PacI  (288)
1 site
T T A A T T A A A A T T A A T T
Bsu36I  (293)
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.
SwaI  (302)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AvrII  (309)
1 site
C C T A G G G G A T C C
PspXI  (320)
1 site
V C T C G A G B B G A G C T C V
BamHI  (1974)
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.
SfiI  (2434)
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.
PaqCI  (2679)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
ScaI  (3424)
1 site
A G T A C T T C A T G A
BstZ17I  (4463)
1 site
G T A T A C C A T A T G
FseI  (5624)
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.
SacII  (5628)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
StuI  (5900)
1 site
A G G C C T T C C G G A
sopA
7457 .. 8632  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
sopA
7457 .. 8632  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
trfA
2080 .. 3228  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds to and activates oriV
trfA
2080 .. 3228  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds to and activates oriV
lacI
10,985 .. 12,067  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
10,985 .. 12,067  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
araC
831 .. 1709  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
araC
831 .. 1709  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
repE
6123 .. 6878  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
repE
6123 .. 6878  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
CmR
3396 .. 4055  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3396 .. 4055  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
oriV
5099 .. 5713  =  615 bp
origin of replication for the bacterial F plasmid
oriV
5099 .. 5713  =  615 bp
origin of replication for the bacterial F plasmid
sopC
9676 .. 10,149  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
sopC
9676 .. 10,149  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
cos
10,409 .. 10,807  =  399 bp
lambda cos site; allows packaging into phage lambda particles
cos
10,409 .. 10,807  =  399 bp
lambda cos site; allows packaging into phage lambda particles
incC
6881 .. 7131  =  251 bp
incompatibility region of the bacterial F plasmid
incC
6881 .. 7131  =  251 bp
incompatibility region of the bacterial F plasmid
MCS
70 .. 314  =  245 bp
multiple cloning site
MCS<