pMCSG23

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

Sequence Author: Midwest Center for Structural Genomics

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: Structural Genomics Vectors | More Plasmid Sets
No matches
T7 promoter EcoNI (4944) AclI (4816) MluI (4476) ApaI (4273) PspOMI (4269) HpaI (3974) BsmBI - Esp3I (3861) PluTI (3841) SfoI (3839) NarI * (3838) KasI (3837) XbaI (3556) SgrAI (2883) NspI (2860) PciI (2856) BmtI (2760) NheI (2756) BspHI (2709) RBS ATG 6xHis AscI (125) PstI - SbfI (135) AflII (163) BsrGI (190) T7 promoter RBS NdeI (298) ATG 6xHis BsaI (405) BspEI (461) BsiWI (632) PsiI (694) BmgBI (884) BsmI (962) AlwNI (987) BsaBI (1017) AfeI (1252) EcoRV (1451) SspI (1487) AvaI - BsoBI - PaeR7I - PspXI - XhoI (1544) PacI (1619) AvrII (1623) EcoO109I (1668) Bsu36I (1707) AgeI (1756) DrdI - PflFI - Tth111I (1816) NgoMIV (2009) NaeI (2011) BglI (2033) pMCSG23 4971 bp
EcoNI  (4944)
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.
AclI  (4816)
1 site
A A C G T T T T G C A A
MluI  (4476)
1 site
A C G C G T T G C G C A
ApaI  (4273)
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  (4269)
1 site
G G G C C C C C C G G G
HpaI  (3974)
1 site
G T T A A C C A A T T G
BsmBI  (3861)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (3861)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
PluTI  (3841)
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  (3839)
1 site
G G C G C C C C G C G G
NarI  (3838)
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  (3837)
1 site
G G C G C C C C G C G G
XbaI  (3556)
1 site
T C T A G A A G A T C T
SgrAI  (2883)
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.
NspI  (2860)
1 site
R C A T G Y Y G T A C R
PciI  (2856)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BmtI  (2760)
1 site
G C T A G C C G A T C G
NheI  (2756)
1 site
G C T A G C C G A T C G
BspHI  (2709)
1 site
T C A T G A A G T A C T
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.
BsaI  (405)
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.
BspEI  (461)
1 site
T C C G G A A G G C C T
BsiWI  (632)
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.
PsiI  (694)
1 site
T T A T A A A A T A T T
BmgBI  (884)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BsmI  (962)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
AlwNI  (987)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BsaBI  (1017)
1 site
G A T N N N N A T C C T A N N N N T A G
AfeI  (1252)
1 site
A G C G C T T C G C G A
EcoRV  (1451)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
SspI  (1487)
1 site
A A T A T T T T A T A A
AvaI  (1544)
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  (1544)
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  (1544)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1544)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1544)
1 site
C T C G A G G A G C T C
PacI  (1619)
1 site
T T A A T T A A A A T T A A T T
AvrII  (1623)
1 site
C C T A G G G G A T C C
EcoO109I  (1668)
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  (1707)
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.
AgeI  (1756)
1 site
A C C G G T T G G C C A
DrdI  (1816)
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.
PflFI  (1816)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1816)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
NgoMIV  (2009)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (2011)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
BglI  (2033)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
ATG
300 .. 302  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
300 .. 302  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
303 .. 320  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
303 .. 320  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
MBP
345 .. 1442  =  1098 bp
366 amino acids  =  40.2 kDa
Product: maltose binding protein from E. coli
This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol.
MBP
345 .. 1442  =  1098 bp
366 amino acids  =  40.2 kDa
Product: maltose binding protein from E. coli
This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol.
TEV site
1464 .. 1484  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
1464 .. 1484  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
lacI
3749 .. 4831  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
3749 .. 4831  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalact