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pRSF-1b

Bacterial vector with an RSF 1030 origin for the regulated expression of N-terminally 6xHis-tagged proteins.

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pRSF-1b.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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PfoI (3606) AclI (3514) BstAPI (3498) MluI (3174) BclI * (3160) BstEII (2992) NmeAIII (2974) ApaI (2971) PspOMI (2967) HpaI (2672) PluTI (2539) SfoI (2537) NarI * (2536) KasI (2535) AcuI (2314) XbaI (2254) BtgI - NcoI (69) BsaAI - PmlI (94) Acc65I (98) KpnI (102) BstBI (107) PshAI (126) BspEI * (130) BamHI (133) MfeI (139) Eco53kI (148) SacI (150) BsrGI - TatI (153) BfuAI - BspMI (160) AscI (161) PstI - SbfI (171) SalI (173) HindIII (179) EagI - NotI (186) PaeR7I - PspXI - XhoI (194) PacI (269) AvrII (273) EcoO109I (318) Bsu36I (357) DrdI (477) BsaI (485) XmnI (573) Bpu10I (942) TspMI - XmaI (1086) SmaI (1088) BspDI - ClaI (1269) NruI (1305) SphI (1494) AfeI (1498) BspQI - SapI (1506) PciI (1622) pRSF-1b 3669 bp
PfoI  (3606)
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  (3514)
1 site
A A C G T T T T G C A A
BstAPI  (3498)
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  (3174)
1 site
A C G C G T T G C G C A
BclI  (3160)
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  (2992)
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.
NmeAIII  (2974)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
ApaI  (2971)
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  (2967)
1 site
G G G C C C C C C G G G
HpaI  (2672)
1 site
G T T A A C C A A T T G
PluTI  (2539)
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  (2537)
1 site
G G C G C C C C G C G G
NarI  (2536)
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  (2535)
1 site
G G C G C C C C G C G G
AcuI  (2314)
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  (2254)
1 site
T C T A G A A G A T C T
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
BsaAI  (94)
1 site
Y A C G T R R T G C A Y
PmlI  (94)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
Acc65I  (98)
1 site
G G T A C C C C A T G G
KpnI  (102)
1 site
G G T A C C C C A T G G
BstBI  (107)
1 site
T T C G A A A A G C T T
PshAI  (126)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BspEI  (130)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
BamHI  (133)
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.
MfeI  (139)
1 site
C A A T T G G T T A A C
Eco53kI  (148)
1 site
G A G C T C C T C G A G
SacI  (150)
1 site
G A G C T C C T C G A G
BsrGI  (153)
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  (153)
1 site
W G T A C W W C A T G W
BfuAI  (160)
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  (160)
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  (161)
1 site
G G C G C G C C C C G C G C G G
PstI  (171)
1 site
C T G C A G G A C G T C
SbfI  (171)
1 site
C C T G C A G G G G A C G T C C
SalI  (173)
1 site
G T C G A C C A G C T G
HindIII  (179)
1 site
A A G C T T T T C G