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pMCSG69 (linearized)

Linearized compact bacterial vector for ligation-independent cloning (LIC), with an MBP-TVMV-6xHis-TEV leader plus tRNA genes for rare Arg and Ile codons.

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pMCSG69 (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Midwest Center for Structural Genomics
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AvaI - BsoBI - PaeR7I - PspXI - XhoI (5902) ScaI (5049) FspI (4791) BglI (4689) AhdI (4569) DrdI (3784) PciI (3676) BspQI - SapI (3560) BstZ17I (3447) PshAI (3421) BsmBI (3191) HpaI (3082) EcoRV (3026) ApaI (2787) PspOMI (2783) BstEII (2757) BmeT110I (5903) EagI - NotI (5910) HindIII (5917) SalI (5923) Eco53kI (5932) SacI (5934) EcoRI (5936) BamHI (5942) End (5948) Start (0) Acc65I (25) KpnI (29) BsrGI (102) BsmI (605) BmgBI (681) BglII (860) BsiWI (929) NdeI (1223) XbaI (1261) BspDI * - ClaI * (1330) AgeI (1372) AscI (1589) SbfI (1600) BstBI (1605) MreI (1737) EcoNI (2111) PflMI (2158) MluI (2576) pMCSG69 5948 bp
AvaI  (5902)
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  (5902)
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  (5902)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5902)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5902)
1 site
C T C G A G G A G C T C
ScaI  (5049)
1 site
A G T A C T T C A T G A
FspI  (4791)
1 site
T G C G C A A C G C G T
BglI  (4689)
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.
AhdI  (4569)
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.
DrdI  (3784)
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.
PciI  (3676)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3560)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3560)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
BstZ17I  (3447)
1 site
G T A T A C C A T A T G
PshAI  (3421)
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.
BsmBI  (3191)
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.
HpaI  (3082)
1 site
G T T A A C C A A T T G
EcoRV  (3026)
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.
ApaI  (2787)
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  (2783)
1 site
G G G C C C C C C G G G
BstEII  (2757)
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.
BmeT110I  (5903)
1 site
C Y C G R G G R G C Y C
EagI  (5910)
1 site
C G G C C G G C C G G C
NotI  (5910)
1 site
G C G G C C G C C G C C G G C G
HindIII  (5917)
1 site
A A G C T T T T C G A A
SalI  (5923)
1 site
G T C G A C C A G C T G
Eco53kI  (5932)
1 site
G A G C T C C T C G A G
SacI  (5934)
1 site
G A G C T C C T C G A G
EcoRI  (5936)
1 site
G A A T T C C T T A A G
BamHI  (5942)
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.
End  (5948)
0 sites
Start  (0)
0 sites
Acc65I  (25)
1 site
G G T A C C C C A T G G
KpnI  (29)
1 site
G G T A C C C C A T G G
BsrGI  (102)
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.
BsmI  (605)
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.
BmgBI  (681)
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.
BglII  (860)
1 site
A G A T C T T C T A G A
BsiWI  (929)
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.
NdeI  (1223)
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.
XbaI  (1261)
1 site
T C T A G A A G A T C T
BspDI  (1330)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1330)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AgeI  (1372)
1 site
A C C G G T T G G C C A
AscI  (1589)
1 site
G G C G C G C C C C G C G C G G
SbfI  (1600)
1 site
C C T G C A G G G G A C G T C C
BstBI  (1605)
1 site
T T C G A A A A G C T T
MreI  (1737)
1 site
C G C C G G C G G C G G C C G C
EcoNI  (2111)
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.
PflMI  (2158)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
MluI  (2576)
1 site
A C G C G T T G C G C A
MBP
124 .. 1224  =  1101 bp
367 amino acids  =  40.3 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
124 .. 1224  =  1101 bp
367 amino acids  =  40.3 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.
lacI
2226 .. 3308  =  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
2226 .. 3308  =  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).
AmpR
4496 .. 5356  =  861 bp
286 amino acids&#