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Plasmid Files

pCOLADuet™-1

Bacterial vector with a ColA origin for the co-expression of two genes.

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pCOLADuet-1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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ACYCDuetUP1 Primer (3660 .. 3678) PfoI (3656) AclI (3564) BstAPI (3548) ApaLI (3244) AflIII - MluI (3224) BclI * (3210) BstEII (3042) ApaI (3021) PspOMI (3017) HpaI (2722) PluTI (2589) SfoI (2587) NarI * (2586) KasI (2585) AcuI (2364) XbaI (2304) SexAI * (2163) BmgBI (1908) BtgI - NcoI (69) BamHI (106) EcoRI (112) Eco53kI (120) SacI (122) BfuAI - BspMI (124) 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) NgoMIV (324) NaeI (326) FseI (328) AsiSI (337) ZraI (344) AatII (346) Acc65I (348) KpnI (352) PaeR7I - PspXI - XhoI (354) PacI (429) AvrII (433) BlpI (451) T7 Terminator Primer (448 .. 466) EcoO109I (478) Bsu36I (517) AgeI (566) DrdI - PflFI - Tth111I (637) BsaI (645) XmnI (733) Bpu10I (1102) TspMI - XmaI (1246) SmaI (1248) BspDI - ClaI (1429) NruI (1465) NheI - NspI - SphI (1654) BmtI (1658) pCOLADuet™-1 3719 bp
PfoI  (3656)
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  (3564)
1 site
A A C G T T T T G C A A
BstAPI  (3548)
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.
ApaLI  (3244)
1 site
G T G C A C C A C G T G
AflIII  (3224)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (3224)
1 site
A C G C G T T G C G C A
BclI  (3210)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (3042)
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.
ApaI  (3021)
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  (3017)
1 site
G G G C C C C C C G G G
HpaI  (2722)
1 site
G T T A A C C A A T T G
PluTI  (2589)
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  (2587)
1 site
G G C G C C C C G C G G
NarI  (2586)
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  (2585)
1 site
G G C G C C C C G C G G
AcuI  (2364)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
XbaI  (2304)
1 site
T C T A G A A G A T C T
SexAI  (2163)
1 site
A C