pETcoco-2

Bacterial vector with an ampicillin 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 PciI (10,151) NcoI (9856) BsaI (9479) SpeI (9391) BseRI (8369) SnaBI (8300) PflFI - Tth111I (7940) KpnI (7880) Acc65I (7876) SbfI (6694) BsrGI (6449) FspAI (6421) 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) EcoRI (2050) SfiI (2434) PaqCI (2679) ScaI (3999) FseI (5567) SacII (5571) StuI (5843) pETcoco™-2 12,417 bp
PciI  (10,151)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NcoI  (9856)
1 site
C C A T G G G G T A C C
BsaI  (9479)
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  (9391)
1 site
A C T A G T T G A T C A
BseRI  (8369)
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  (8300)
1 site
T A C G T A A T G C A T
PflFI  (7940)
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  (7940)
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  (7880)
1 site
G G T A C C C C A T G G
Acc65I  (7876)
1 site
G G T A C C C C A T G G
SbfI  (6694)
1 site
C C T G C A G G G G A C G T C C
BsrGI  (6449)
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  (6421)
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