pMCS5

E. coli cloning vector with an extensive multiple cloning site containing 59 unique restriction sites.

Sequence Author: MoBiTec

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PstI - SbfI (211) SphI (205) BfuAI - BspMI (200) HindIII (195) RsrII (189) AvrII (182) I-SceI (180) PsiI (2973) DraIII (2848) BtgZI (2840) XmnI (2295) ScaI (2176) TatI (2174) NmeAIII (1844) BpmI (1766) BsaI (1757) AhdI (1696) SalI (213) AccI (214) XbaI (219) BamHI (225) PmlI (233) TspMI - XmaI (237) SmaI - SrfI (239) PmeI (247) MfeI (252) BstBI (259) BsiWI (264) NruI (272) PaeR7I - PspXI - XhoI (276) SnaBI (284) Acc65I (288) KpnI (292) Eco53kI (296) SacI (298) EcoRI (300) EagI - NotI (307) SacII (313) PspOMI (314) ApaI (318) NdeI (320) SfiI (322) AgeI - SgrAI (329) FseI (341) BglII (344) NheI (350) BmtI (354) SpeI (356) KasI (362) NarI (363) SfoI (364) PluTI (366) MluI (368) EcoRV (376) NsiI (384) HpaI (388) BspDI * - ClaI * (393) NcoI (398) AscI - BssHII (404) PacI (415) SwaI (422) lac operator BspQI - SapI (687) PciI (803) BseYI (1107) PspFI (1111) AlwNI (1219) pMCS5 3081 bp
PstI  (211)
1 site
C T G C A G G A C G T C
SbfI  (211)
1 site
C C T G C A G G G G A C G T C C
SphI  (205)
1 site
G C A T G C C G T A C G
BfuAI  (200)
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  (200)
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.
HindIII  (195)
1 site
A A G C T T T T C G A A
RsrII  (189)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
AvrII  (182)
1 site
C C T A G G G G A T C C
I-SceI  (180)
1 site
T A G G G A T A A C A G G G T A A T A T C C C T A T T G T C C C A T T A

I-SceI is a homing endonuclease that can recognize a variety of similar recognition sequences.
After cleavage, I-SceI can remain bound to DNA and alter its electrophoretic mobility.
PsiI  (2973)
1 site
T T A T A A A A T A T T
DraIII  (2848)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BtgZI  (2840)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
XmnI  (2295)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2176)
1 site
A G T A C T T C A T G A
TatI  (2174)
1 site
W G T A C W W C A T G W
NmeAIII  (1844)
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).
BpmI  (1766)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BsaI  (1757)
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.
AhdI  (1696)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
SalI  (213)
1 site
G T C G A C C A G C T G
AccI  (214)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
XbaI  (219)
1 site
T C T A G A A G A T C T
BamHI  (225)
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.
PmlI  (233)
1 site
C A C G T G G T G C A C
TspMI  (237)
1 site
C C C G G G G G G C C C
XmaI  (237)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (239)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (239)
1 site
G C C C G G G C C G G G C C C G
PmeI  (247)
1 site
G T T T A A A C C A A A T T T G
MfeI  (252)
1 site
C A A T T G G T T A A C
BstBI  (259)
1 site
T T C G A A A A G C T T
BsiWI  (264)
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.
NruI  (272)
1 site
T C G C G A A G C G C T
PaeR7I  (276)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (276)
1 site
V C T C G A G B B G A G C T C V
XhoI  (276)
1 site
C T C G A G G A G C T C
SnaBI  (284)
1 site
T A C G T A A T G C A T
Acc65I  (288)
1 site
G G T A C C C C A T G G
KpnI  (292)
1 site
G G T A C C C C A T G G
Eco53kI  (296)
1 site
G A G C T C C T C G A G
SacI  (298)
1 site
G A G C T C C T C G A G
EcoRI  (300)
1 site
G A A T T C C T T A A G
EagI  (307)
1 site
C G G C C G G C C G G C
NotI  (307)
1 site
G C G G C C G C C G C C G G C G
SacII  (313)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
PspOMI  (314)
1 site
G G G C C C C C C G G G
ApaI  (318)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
NdeI  (320)
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.
SfiI  (322)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AgeI  (329)
1 site
A C C G G T T G G C C A
SgrAI  (329)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
FseI  (341)
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.
BglII  (344)
1 site
A G A T C T T C T A G A
NheI  (350)
1 site
G C T A G C C G A T C G
BmtI  (354)
1 site
G C T A G C C G A T C G
SpeI  (356)
1 site
A C T A G T T G