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pMCSG22

Bacterial coexpression vector with a 6xHis-S- loop-TEV leader and a CloDF13 origin, for high-throughput purification of recombinant proteins.

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pMCSG22.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Midwest Center for Structural Genomics
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EcoNI (3879) PfoI (3843) AclI (3751) BstAPI (3735) MluI (3411) ApaI (3208) PspOMI (3204) HpaI (2909) BsmBI (2796) PluTI (2776) SfoI (2774) NarI * (2773) KasI (2772) XbaI (2491) SgrAI (1818) NcoI (69) AscI (125) PstI - SbfI (135) AflII (163) BsrGI (190) NdeI (298) BglII (332) Acc65I (393) KpnI (397) SspI (422) AvaI - BsoBI - PaeR7I - PspXI - XhoI (479) BmeT110I (480) PacI (554) AvrII (558) EcoO109I (603) Bsu36I (642) DrdI - PflFI - Tth111I (751) BsaAI (840) NgoMIV (944) NaeI (946) BglI (968) PvuI (1218) BspHI (1644) NheI (1691) BmtI (1695) PciI (1791) NspI (1795) pMCSG22 3906 bp
EcoNI  (3879)
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  (3843)
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  (3751)
1 site
A A C G T T T T G C A A
BstAPI  (3735)
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  (3411)
1 site
A C G C G T T G C G C A
ApaI  (3208)
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  (3204)
1 site
G G G C C C C C C G G G
HpaI  (2909)
1 site
G T T A A C C A A T T G
BsmBI  (2796)
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  (2776)
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  (2774)
1 site
G G C G C C C C G C G G
NarI  (2773)
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  (2772)
1 site
G G C G C C C C G C G G
XbaI  (2491)
1 site
T C T A G A A G A T C T
SgrAI  (1818)
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.
NcoI  (69)
1 site
C C A T G G G G T A C C
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
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.
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  (332)
1 site
A G A T C T T C T A G A
Acc65I  (393)
1 site
G G T A C C C C A T G G
KpnI  (397)
1 site
G G T A C C C C A T G G
SspI  (422)
1 site
A A T A T T T T A T A A
AvaI  (479)
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  (479)
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  (479)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (479)
1 site
V C T C G A G B B G A G C T C V
XhoI  (479)
1 site
C T C G A G G A G C T C
BmeT110I  (480)
1 site
C Y C G R G G R G C Y C
PacI  (554)
1 site
T T A A T T A A A A T T A A T T
AvrII  (558)
1 site
C C T A G G G G A T C C
EcoO109I  (603)
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  (642)
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.
DrdI  (751)
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.