Resources
Plasmid Files

pMCSG27 (linearized)

Linearized bacterial vector for ligation-independent cloning (LIC), with a C-terminal 10xHis-TVMV-MBP cassette.

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

pMCSG27 (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Midwest Center for Structural Genomics
Download Free Trial Get SnapGene Viewer


AvaI - BsoBI - PaeR7I - PspXI - XhoI (5156) ScaI (4303) PstI (4068) AhdI (3823) PciI (2930) BspQI - SapI (2814) BstZ17I (2701) PflFI - Tth111I (2675) Bpu10I (2036) PpuMI (1936) FspAI (1911) PshAI (1674) BmeT110I (5157) EagI - NotI (5164) HindIII (5171) SalI (5177) Eco53kI (5186) SacI (5188) EcoRI (5190) BamHI (5196) NcoI (5345) BsmI (5692) BmgBI (5768) BglII (5947) PsiI (5958) BsiWI (6016) End (6351) Start (0) XbaI (37) BspDI * - ClaI * (106) SgrAI (148) SphI (304) EcoNI (364) PflMI (411) MluI (829) BstEII (1010) PspOMI (1036) ApaI (1040) EcoRV (1279) HpaI (1335) pMCSG27 6351 bp
AvaI  (5156)
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  (5156)
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  (5156)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5156)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5156)
1 site
C T C G A G G A G C T C
ScaI  (4303)
1 site
A G T A C T T C A T G A
PstI  (4068)
1 site
C T G C A G G A C G T C
AhdI  (3823)
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.
PciI  (2930)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2814)
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  (2814)
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  (2701)
1 site
G T A T A C C A T A T G
PflFI  (2675)
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  (2675)
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.
Bpu10I  (2036)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
PpuMI  (1936)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
FspAI  (1911)
1 site
R T G C G C A Y Y A C G C G T R
PshAI  (1674)
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.
BmeT110I  (5157)
1 site
C Y C G R G G R G C Y C
EagI  (5164)
1 site
C G G C C G G C C G G C
NotI  (5164)
1 site
G C G G C C G C C G C C G G C G
HindIII  (5171)
1 site
A A G C T T T T C G A A
SalI  (5177)
1 site
G T C G A C C A G C T G
Eco53kI  (5186)
1 site
G A G C T C C T C G A G
SacI  (5188)
1 site
G A G C T C C T C G A G
EcoRI  (5190)
1 site
G A A T T C C T T A A G
BamHI  (5196)
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.
NcoI  (5345)
1 site
C C A T G G G G T A C C
BsmI  (5692)
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  (5768)
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  (5947)
1 site
A G A T C T T C T A G A
PsiI  (5958)
1 site
T T A T A A A A T A T T
BsiWI  (6016)
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.
End  (6351)
0 sites
Start  (0)
0 sites
XbaI  (37)
1 site
T C T A G A A G A T C T
BspDI  (106)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (106)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SgrAI  (148)
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.
SphI  (304)
1 site
G C A T G C C G T A C G
EcoNI  (364)
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  (411)
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  (829)
1 site
A C G C G T T G C G C A
BstEII  (1010)
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.
PspOMI  (1036)
1 site
G G G C C C C C C G G G
ApaI  (1040)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoRV  (1279)
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.
HpaI  (1335)
1 site
G T T A A C C A A T T G
MBP
5211 .. 6308  =  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
5211 .. 6308  =  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.
lacI
479 .. 1561  =  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
479 .. 1561  =  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
3750 .. 4610  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3750 .. 4541  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3750 .. 4610  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   4542 .. 4610  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3750 .. 4610  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2991 .. 3579  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2991 .. 3579  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
2370 .. 2561  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
2370 .. 2561  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
4611 .. 4714  =  104 bp
AmpR promoter
4611 .. 4714  =  104 bp
lacI promoter
401 .. 478  =  78 bp
lacI promoter
401 .. 478  =  78 bp
T7 terminator
5024 .. 5071  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
5024 .. 5071  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
10xHis
6330 .. 6359  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
10xHis
6330 .. 6359  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
lac operator
45 .. 69  =  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
45 .. 69  =  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).
TVMV site
6309 .. 6329  =  21 bp
7 amino acids  =  866.0 Da
Product:
tobacco vein mottling virus (TVMV) NIa protease recognition and cleavage site
TVMV site
6309 .. 6329  =  21 bp
7 amino acids  =  866.0 Da
Product:
tobacco vein mottling virus (TVMV) NIa protease recognition and cleavage site
T7 promoter
70 .. 88  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
70 .. 88  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
5138 .. 5155  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5138 .. 5155  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
9 .. 14  =  6 bp
ribosome binding site
RBS
9 .. 14  =  6 bp
ribosome binding site
ORF:  1 .. 729  =  729 bp
ORF:  242 amino acids  =  25.4 kDa  (no start codon)
ORF:  1318 .. 1581  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3880 .. 4146  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  602 .. 1561  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  1617 .. 1973  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  2337 .. 2561  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  177 .. 440  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  1344 .. 1595  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  3750 .. 4610  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  5202 .. 6362  =  1161 bp
ORF:  386 amino acids  =  42.7 kDa  (no start codon)
ORF:  1970 .. 2338  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  52 .. 303  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
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 © 2018 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter