pMCSG68

Compact bacterial vector encoding tRNA genes for rare Arg and Ile codons, with a 6xHis-Strep-Tag® II-TEV leader 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
AvaI - BsoBI - PaeR7I - PspXI - XhoI (158) BlpI (80) T7 terminator BspEI (2) ScaI (4137) PvuI (4027) FspI (3879) BglI (3777) AhdI (3657) AlwNI (3180) DrdI (2872) PciI (2764) BspQI - SapI (2648) BstZ17I (2535) BsaAI (2516) PshAI (2509) EagI - NotI (166) HindIII (173) SalI (179) Eco53kI (188) SacI (190) EcoRI (192) BamHI (198) SspI (219) TEV site Acc65I (244) KpnI (248) Strep-Tag II BglII (275) 6xHis ATG NdeI (311) XbaI (349) lac operator T7 promoter BspDI * - ClaI * (418) AgeI (460) AscI (677) SbfI (688) BfuAI - BspMI (706) MreI (825) AfeI (911) NcoI (1095) PsiI (1110) EcoNI (1199) PfoI (1231) PflMI (1246) BstAPI (1347) MluI (1664) BclI * (1678) BstEII (1845) PspOMI (1871) ApaI (1875) EcoRV (2114) HpaI (2170) BsmBI - Esp3I (2279) pMCSG68 4832 bp
AvaI  (158)
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  (158)
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  (158)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site
V C T C G A G B B G A G C T C V
XhoI  (158)
1 site
C T C G A G G A G C T C
BlpI  (80)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BspEI  (2)
1 site
T C C G G A A G G C C T
ScaI  (4137)
1 site
A G T A C T T C A T G A
PvuI  (4027)
1 site
C G A T C G G C T A G C
FspI  (3879)
1 site
T G C G C A A C G C G T
BglI  (3777)
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  (3657)
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.
AlwNI  (3180)
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.
DrdI  (2872)
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  (2764)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2648)
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  (2648)
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  (2535)
1 site
G T A T A C C A T A T G
BsaAI  (2516)
1 site
Y A C G T R R T G C A Y
PshAI  (2509)
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.
EagI  (166)
1 site
C G G C C G G C C G G C
NotI  (166)
1 site
G C G G C C G C C G C C G G C G
HindIII  (173)
1 site
A A G C T T T T C G A A
SalI  (179)
1 site
G T C G A C C A G C T G
Eco53kI  (188)
1 site
G A G C T C C T C G A G
SacI  (190)
1 site
G A G C T C C T C G A G
EcoRI  (192)
1 site
G A A T T C C T T A A G
BamHI  (198)
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.
SspI  (219)
1 site
A A T A T T T T A T A A
Acc65I  (244)
1 site
G G T A C C C C A T G G
KpnI  (248)
1 site
G G T A C C C C A T G G
BglII  (275)
1 site
A G A T C T T C T A G A
NdeI  (311)
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  (349)
1 site
T C T A G A A G A T C T
BspDI  (418)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (418)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AgeI  (460)
1 site
A C C G G T T G G C C A
AscI  (677)
1 site
G G C G C G C C C C G C G C G G
SbfI  (688)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (706)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (706)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
MreI  (825)
1 site
C G C C G G C G G C G G C C G C
AfeI  (911)
1 site
A G C G C T T C G C G A
NcoI  (1095)
1 site
C C A T G G G G T A C C
PsiI  (1110)
1 site
T T A T A A A A T A T T
EcoNI  (1199)
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  (1231)
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.
PflMI  (1246)
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.
BstAPI  (1347)
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  (1664)
1 site
A C G C G T T G C G C A
BclI  (1678)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (1845)
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  (1871)
1 site
G G G C C C C C C G G G
ApaI  (1875)
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  (2114)
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  (2170)
1 site
G T T A A C C A A T T G
BsmBI  (2279)
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  (2279)
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.
lacI
1314 .. 2396  =  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
1314 .. 2396  =  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
3584 .. 4444  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   3584 .. 4375  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3584 .. 4444  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   4376 .. 4444  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3584 .. 4444  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2825 .. 3413  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2825 .. 3413  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
4445 .. 4548  =  104 bp
AmpR promoter
4445 .. 4548  =  104 bp
TEV site
223 .. 243  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
223 .. 243  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
Strep-Tag II
250 .. 273  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
Strep-Tag II
250 .. 273  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
6xHis
292 .. 309  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
292 .. 309  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
310 .. 312  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
310 .. 312  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
lacI promoter
1236 .. 1313  =  78 bp
lacI promoter
1236 .. 1313  =  78 bp
argU
675 .. 751  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
argU
675 .. 751  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
ileX
1021 .. 1096  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
ileX
1021 .. 1096  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
357 .. 381  =  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
357 .. 381  =  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).
T7 promoter
382 .. 400  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
382 .. 400  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
321 .. 326  =  6 bp
ribosome binding site
RBS
321 .. 326  =  6 bp
ribosome binding site
ORF:  265 .. 525  =  261 bp
ORF:  86 amino acids  =  9.8 kDa
ORF:  1097 .. 1564  =  468 bp
ORF:  155 amino acids  =  16.8 kDa
ORF:  2153 .. 2416  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  810 .. 1064  =  255 bp
ORF:  84 amino acids  =  8.5 kDa
ORF:  1437 .. 2396  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  3714 .. 3980  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3584 .. 4444  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2179 .. 2430  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
Click here to try SnapGene

Download pMCSG68.dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps