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pCDFDuet™-1

Bacterial vector with a CloDF13 (CDF) origin for the co-expression of two genes.

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pCDFDuet-1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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NcoI (69) EcoNI (3754) ACYCDuetUP1 Primer (3722 .. 3740) PfoI (3718) AclI (3626) BstAPI (3610) MluI (3286) ApaI (3083) PspOMI (3079) HpaI (2784) BsmBI (2671) PluTI (2651) SfoI (2649) NarI * (2648) KasI (2647) XbaI (2366) BamHI (106) EcoRI (112) Eco53kI (120) SacI (122) AscI (125) PstI - SbfI (135) SalI (137) HindIII (143) NotI (150) AflII (163) DuetUP2 Primer (189 .. 208) BsrGI - TatI (190) DuetDOWN1 Primer (189 .. 208) NdeI (298) BglII (305) MfeI (311) EcoRV (319) FseI (328) AsiSI (337) ZraI (344) AatII (346) Acc65I (348) KpnI (352) AvaI - BsoBI - PaeR7I - PspXI - XhoI (354) BmeT110I (355) PacI (429) AvrII (433) T7 Terminator Primer (448 .. 466) EcoO109I (478) Bsu36I (517) AgeI (566) DrdI - PflFI - Tth111I (626) BsaAI (715) BglI (843) SspI (1488) BspHI (1519) NheI (1566) BmtI (1570) PciI (1666) NspI (1670) SgrAI (1693) pCDFDuet™-1 3781 bp
NcoI  (69)
1 site
C C A T G G G G T A C C
EcoNI  (3754)
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.
PfoI  (3718)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
AclI  (3626)
1 site
A A C G T T T T G C A A
BstAPI  (3610)
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.
MluI  (3286)
1 site
A C G C G T T G C G C A
ApaI  (3083)
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  (3079)
1 site
G G G C C C C C C G G G
HpaI  (2784)
1 site
G T T A A C C A A T T G
BsmBI  (2671)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
PluTI  (2651)
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  (2649)
1 site
G G C G C C C C G C G G
NarI  (2648)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (2647)
1 site
G G C G C C C C G C G G
XbaI  (2366)
1 site
T C T A G A A G A T C T
BamHI  (106)
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  (112)
1 site
G A A T T C C T T A A G
Eco53kI  (120)
1 site
G A G C T C C T C G A G
SacI  (122)
1 site
G A G C T C C T C G A G
AscI  (125)
1 site
G G C G C G C C C C G C G C G G
PstI  (135)
1 site
C T G C A G G A C G T C
SbfI  (135)
1 site
C C T G C A G G G G A C G T C C
SalI  (137)
1 site
G T C G A C C A G C T G
HindIII  (143)
1 site
A A G C T T T T C G A A
NotI  (150)
1 site
G C G G C C G C C G C C G G C G
AflII  (163)
1 site
C T T A A G G A A T T C
BsrGI  (190)
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.
TatI  (190)
1 site
W G T A C W W C A T G W
NdeI  (298)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BglII  (305)
1 site
A G A T C T T C T A G A
MfeI  (311)
1 site
C A A T T G G T T A A C
EcoRV  (319)
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.
FseI  (328)
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.
AsiSI  (337)
1 site
G C G A T C G C C G C T A G C G
ZraI  (344)
1 site
G A C G T C C T G C A G
AatII  (346)
1 site
G A C G T C C T G C A G
Acc65I  (348)
1 site
G G T A C C C C A T G G
KpnI  (352)
1 site
G G T A C C C C A T G G
AvaI  (354)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (354)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (354)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (354)
1 site
V C T C G A G B B G A G C T C V
XhoI  (354)
1 site
C T C G A G G A G C T C
BmeT110I  (355)
1 site
C Y C G R G G R G C Y C
PacI  (429)
1 site
T T A A T T A A A A T T A A T T
AvrII  (433)
1 site
C C T A G G G G A T C C
EcoO109I  (478)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
Bsu36I  (517)
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.
AgeI  (566)
1 site
A C C G G T T G G C C A
DrdI  (626)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PflFI  (626)
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  (626)
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.
BsaAI  (715)
1 site
Y A C G T R R T G C A Y
BglI  (843)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
SspI  (1488)
1 site
A A T A T T T T A T A A
BspHI  (1519)
1 site
T C A T G A A G T A C T
NheI  (1566)
1 site
G C T A G C C G A T C G
BmtI  (1570)
1 site
G C T A G C C G A T C G
PciI  (1666)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (1670)
1 site
R C A <