Resources
Plasmid Files

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.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pMCSG69 (linearized) Sequence and MappMCSG69 (linearized).dna
Map and Sequence File   
Sequence Author:  Midwest Center for Structural Genomics
Download Free Trial Get SnapGene Viewer

 6xHis 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) AvaI - BsoBI - PaeR7I - PspXI - XhoI (5902) BmeT110I (5903) EagI - NotI (5910) HindIII (5917) SalI (5923) Eco53kI (5932) SacI (5934) EcoRI (5936) BamHI (5942) End (5948) Start (0) TEV site Acc65I (25) KpnI (29) 6xHis BsrGI (102) BsmI (605) BmgBI (681) BglII (860) BsiWI (929) NdeI (1223) RBS XbaI (1261) T7 promoter BspDI * - ClaI * (1330) AgeI (1372) AscI (1589) SbfI (1600) BstBI (1605) MreI (1737) EcoNI (2111) PflMI (2158) MluI (2576) pMCSG69 5948 bp
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.
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
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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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  =  31.5 kDa
   Segment 2:  
   4496 .. 5287  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4496 .. 5356  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5288 .. 5356  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4496 .. 5356  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3737 .. 4325  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3737 .. 4325  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
5357 .. 5460  =  104 bp
AmpR promoter
5357 .. 5460  =  104 bp
lacI promoter
2148 .. 2225  =  78 bp
lacI promoter
2148 .. 2225  =  78 bp
argU
1587 .. 1663  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
argU
1587 .. 1663  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
ileX
1933 .. 2008  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
ileX
1933 .. 2008  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
T7 terminator
5770 .. 5817  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
5770 .. 5817  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
1269 .. 1293  =  25 bp
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).
lac operator
1269 .. 1293  =  25 bp
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).
TEV site
4 .. 24  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV site
4 .. 24  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TVMV site
67 .. 87  =  21 bp
7 amino acids  =  866.0 Da
Product:
tobacco vein mottling virus (TVMV) NIa
protease recognition and cleavage site
TVMV site
67 .. 87  =  21 bp
7 amino acids  =  866.0 Da
Product:
tobacco vein mottling virus (TVMV) NIa
protease recognition and cleavage site
T7 promoter
1294 .. 1312  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1294 .. 1312  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
49 .. 66  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
49 .. 66  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5884 .. 5901  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5884 .. 5901  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
1233 .. 1238  =  6 bp
ribosome binding site
RBS
1233 .. 1238  =  6 bp
ribosome binding site
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter