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pRSFDuet™-1

Bacterial vector with an RSF 1030 origin for the co-expression of two genes.

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pRSFDuet-1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  MilliporeSigma (Novagen)
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BtgI - NcoI (69) ACYCDuetUP1 Primer (3770 .. 3788) PfoI (3766) AclI (3674) BstAPI (3658) MluI (3334) BclI * (3320) BstEII (3152) ApaI (3131) PspOMI (3127) HpaI (2832) PluTI (2699) SfoI (2697) NarI * (2696) KasI (2695) AcuI (2474) XbaI (2414) BssS α I (1944) 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) EcoRV (319) NgoMIV (324) NaeI (326) FseI (328) AsiSI - PvuI (337) ZraI (344) AatII (346) Acc65I (348) KpnI (352) PaeR7I - PspXI - XhoI (354) PacI (429) AvrII (433) T7 Terminator Primer (448 .. 466) EcoO109I (478) Bsu36I (517) AgeI (566) DrdI (637) BsaI (645) XmnI (733) Bpu10I (1102) TspMI - XmaI (1246) SmaI (1248) BspDI - ClaI (1429) NruI (1465) SphI (1654) AfeI (1658) BspQI - SapI (1666) PciI (1782) pRSFDuet™-1 3829 bp
BtgI  (69)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (69)
1 site
C C A T G G G G T A C C
PfoI  (3766)
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  (3674)
1 site
A A C G T T T T G C A A
BstAPI  (3658)
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  (3334)
1 site
A C G C G T T G C G C A
BclI  (3320)
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  (3152)
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  (3131)
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  (3127)
1 site
G G G C C C C C C G G G
HpaI  (2832)
1 site
G T T A A C C A A T T G
PluTI  (2699)
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  (2697)
1 site
G G C G C C C C G C G G
NarI  (2696)
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  (2695)
1 site
G G C G C C C C G C G G
AcuI  (2474)
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  (2414)
1 site
T C T A G A A G A T C T
BssSαI  (1944)
1 site
C A C G A G G T G C T C
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
BfuAI  (124)
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  (124)
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.
AscI  (125)
1 site
G