pMCSG31 (linearized)

Linearized bacterial vector for ligation-independent cloning (LIC), with an MBP-TVMV-6xHis-TEV leader and a Tet-inducible TVMV protease cassette.

Sequence Author: Midwest Center for Structural Genomics

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HomePlasmidsStructural Genomics VectorspMCSG31 (linearized)
PflMI (3274) BsgI (2397) AleI (2373) NruI (2115) SnaBI (2072) EcoNI (2043) Bpu10I (1996) AhdI (1752) BpmI (1683) NmeAIII (1605) MfeI (1574) FspI (1529) ScaI (1271) XmnI (1152) BstBI (279) SbfI (240) AbsI (224) SmaI (215) TspMI - XmaI (213) BseRI (3513) AgeI (3678) BsmBI - Esp3I (3819) BbsI (3863) AflIII - MluI (4177) SphI (4298) SgrAI (4446) BspDI * - ClaI * (4490) T7 promoter RBS PfoI * (4855) BsiWI (4889) BglII (4958) BstXI (4984) BssHII (5035) BmgBI (5141) BsrGI (5716) 6xHis Acc65I (5793) KpnI (5797) End (5807) Start (0) Eco53kI (31) SacI (33) EagI - NotI (49) 6xHis pMCSG31 5807 bp
PflMI  (3274)
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.
BsgI  (2397)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AleI  (2373)
1 site		 C A C N N N N G T G G T G N N N N C A C
NruI  (2115)
1 site		 T C G C G A A G C G C T
SnaBI  (2072)
1 site		 T A C G T A A T G C A T
EcoNI  (2043)
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.
Bpu10I  (1996)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
AhdI  (1752)
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.
BpmI  (1683)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
NmeAIII  (1605)
1 site		 G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19
Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
MfeI  (1574)
1 site		 C A A T T G G T T A A C
FspI  (1529)
1 site		 T G C G C A A C G C G T
ScaI  (1271)
1 site		 A G T A C T T C A T G A
XmnI  (1152)
1 site		 G A A N N N N T T C C T T N N N N A A G
BstBI  (279)
1 site		 T T C G A A A A G C T T
SbfI  (240)
1 site		 C C T G C A G G G G A C G T C C
AbsI  (224)
1 site		 C C T C G A G G G G A G C T C C
SmaI  (215)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
TspMI  (213)
1 site		 C C C G G G G G G C C C
XmaI  (213)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
BseRI  (3513)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
AgeI  (3678)
1 site		 A C C G G T T G G C C A
BsmBI  (3819)
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  (3819)
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.
BbsI  (3863)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
AflIII  (4177)
1 site		 A C R Y G T T G Y R C A
Sticky ends from different AflIII sites may not be compatible.
MluI  (4177)
1 site		 A C G C G T T G C G C A
SphI  (4298)
1 site		 G C A T G C C G T A C G
SgrAI  (4446)
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.
BspDI  (4490)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (4490)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
PfoI  (4855)
1 site		 T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BsiWI  (4889)
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.
BglII  (4958)
1 site		 A G A T C T T C T A G A
BstXI  (4984)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
BssHII  (5035)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
BmgBI  (5141)
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.
BsrGI  (5716)
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.
Acc65I  (5793)
1 site		 G G T A C C C C A T G G
KpnI  (5797)
1 site		 G G T A C C C C A T G G
End  (5807)
0 sites
Start  (0)
0 sites
Eco53kI  (31)
1 site		 G A G C T C C T C G A G
SacI  (33)
1 site		 G A G C T C C T C G A G
EagI  (49)
1 site		 C G G C C G G C C G G C
NotI  (49)
1 site		 G C G G C C G C C G C C G G C G
MBP
4599 .. 5699  =  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
4599 .. 5699  =  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.
TVMV site
5736 .. 5756  =  21 bp
7 amino acids  =  865.9 Da
Product:
tobacco vein mottling virus (TVMV) NIa protease recognition and cleavage site
TVMV site
5736 .. 5756  =  21 bp
7 amino acids  =  865.9 Da
Product:
tobacco vein mottling virus (TVMV) NIa protease recognition and cleavage site
6xHis
5757 .. 5774  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5757 .. 5774  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
TEV site
5799 .. 5819  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
5799 .. 5819  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
AmpR
965 .. 1825  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   965 .. 1033  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
965 .. 1825  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   1034 .. 1825  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
965 .. 1825  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TVMV protease
3451 .. 4161  =  711 bp
236 amino acids  =  26.6 kDa
Product: tobacco vein mottling virus NIa protease (Nallamsetty et al., 2004)
TVMV protease
3451 .. 4161  =  711 bp
236 amino acids  =  26.6 kDa
Product: tobacco vein mottling virus NIa protease (Nallamsetty et al., 2004)
TetR
1832 .. 2458  =  627 bp
208 amino acids  =  23.5 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
TetR
1832 .. 2458  =  627 bp
208 amino acids  =  23.5 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
ori
2611 .. 3199  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2611 .. 3199  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
377 .. 835  =  459 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
377 .. 835  =  459 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
861 .. 964  =  104 bp
AmpR promoter
861 .. 964  =  104 bp
rrnB T1 terminator
4194 .. 4280  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4194 .. 4280  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
tetR/tetA promoters
3303 .. 3358  =  56 bp
overlapping promoters for bacterial tetR and tetA
tetR/tetA promoters
3303 .. 3358  =  56 bp
overlapping promoters for bacterial tetR and tetA
T7 terminator
147 .. 194  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
147 .. 194  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
4530 .. 4554  =  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
4530 .. 4554  =  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).
Strep-Tag II
263 .. 286  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
Strep-Tag II
263 .. 286  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
T7 promoter
4511 .. 4529  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4511 .. 4529  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
63 .. 80  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
63 .. 80  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
4585 .. 4590  =  6 bp
ribosome binding site
RBS
4585 .. 4590  =  6 bp
ribosome binding site
tet operator
3309 .. 3327  =  19 bp
bacterial operator O1 for the tetR and tetA genes
tet operator
3309 .. 3327  =  19 bp
bacterial operator O1 for the tetR and tetA genes
tet operator
3339 .. 3357  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
3339 .. 3357  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  3427 .. 4161  =  735 bp
ORF:  244 amino acids  =  27.5 kDa
ORF:  965 .. 1825  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1832 .. 2458  =  627 bp
ORF:  208 amino acids  =  23.5 kDa
ORF:  4028 .. 4273  =  246 bp
ORF:  81 amino acids  =  9.2 kDa
ORF:  4296 .. 4547  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
ORF:  4599 .. 5822  =  1224 bp
ORF:  408 amino acids  =  45.0 kDa
ORF:  4158 .. 4718  =  561 bp
ORF:  186 amino acids  =  20.2 kDa
ORF:  4938 .. 5822  =  885 bp
ORF:  294 amino acids  =  34.1 kDa  (no start codon)
ORF:  2 .. 334  =  333 bp
ORF:  111 amino acids  =  12.2 kDa
ORF:  1429 .. 1695  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1756 .. 2004  =  249 bp
ORF:  82 amino acids  =  8.9 kDa
ORF:  3253 .. 3852  =  600 bp
ORF:  199 amino acids  =  23.2 kDa
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Individual Sequences & Maps

p11
pMCSG19B (linearized)
pMCSG41
pMCSG68 (linearized)
p15Tv-L
pMCSG19C
pMCSG41 (linearized)
pMCSG69
p283
pMCSG19C (linearized)
pMCSG42
pMCSG69 (linearized)
pBEN1-SGC
pMCSG20
pMCSG42 (linearized)
pMCSG7
pBEN1-SGC (linearized)
pMCSG20 (linearized)
pMCSG43
pMCSG7 (linearized)
pDB-HisGST
pMCSG21
pMCSG43 (linearized)
pMCSG70
pDB.GST
pMCSG21 (linearized)
pMCSG46
pMCSG70 (linearized)
pDB.His.MBP
pMCSG22
pMCSG46 (linearized)
pMCSG71
pDB.His.NusA
pMCSG22 (linearized)
pMCSG48
pMCSG71 (linearized)
pDB.His.TRX
pMCSG23
pMCSG48 (linearized)
pMCSG73
pDB.TRX
pMCSG23 (linearized)
pMCSG49
pMCSG73 (linearized)
pEU-GST-FV
pMCSG24
pMCSG49 (linearized)
pMCSG75
pEU-His-FV
pMCSG24 (linearized)
pMCSG50
pMCSG75 (linearized)
pEU-HSBC
pMCSG25
pMCSG50 (linearized)
pMCSG76
pFB-LIC-Bse
pMCSG25 (linearized)
pMCSG51
pMCSG76 (linearized)
pFB-LIC-Bse (linearized)
pMCSG26
pMCSG51 (linearized)
pMCSG77
pH3c-LIC
pMCSG26 (linearized)
pMCSG52
pMCSG77 (linearized)
pH3c-LIC (linearized)
pMCSG27
pMCSG52 (linearized)
pMCSG79
pHM3C-LIC
pMCSG27 (linearized)
pMCSG53
pMCSG79 (linearized)
pHM3C-LIC (linearized)
pMCSG28
pMCSG53 (linearized)
pMCSG8
pLIC-CH
pMCSG28 (linearized)
pMCSG54
pMCSG8 (linearized)
pLIC-CH (linearized)
pMCSG29
pMCSG54 (linearized)
pMCSG81
pLIC-EGFP
pMCSG29 (linearized)
pMCSG55
pMCSG81 (linearized)
pLIC-EGFP (linearized)
pMCSG30
pMCSG55 (linearized)
pMCSG9
pLIC-SGC1
pMCSG30 (linearized)
pMCSG56
pMCSG9 (linearized)
pLIC-SGC1 (linearized)
pMCSG31
pMCSG56 (linearized)
pNH-TrxT
pMCentr1
pMCSG31 (linearized)
pMCSG57
pNH-TrxT (linearized)
pMCentr1 (linearized)
pMCSG32
pMCSG57 (linearized)
pNIC-CH
pMCentr2
pMCSG32 (linearized)
pMCSG58
pNIC-CH (linearized)
pMCentr2 (linearized)
pMCSG33
pMCSG58 (linearized)
pNIC-CTHF
pMCentr3
pMCSG33 (linearized)
pMCSG59
pNIC-CTHF (linearized)
pMCentr3 (linearized)
pMCSG34
pMCSG59 (linearized)
pNIC-Zb
pMCSG10
pMCSG34 (linearized)
pMCSG60
pNIC-Zb (linearized)
pMCSG10 (linearized)
pMCSG34B
pMCSG60 (linearized)
pNIC28-Bsa4
pMCSG11
pMCSG34B (linearized)
pMCSG61
pNIC28-Bsa4 (linearized)
pMCSG11 (linearized)
pMCSG35
pMCSG61 (linearized)
pNYCOMPS-LIC-FH10T+ (N term) - linearized
pMCSG12
pMCSG35 (linearized)
pMCSG62
pNYCOMPS-LIC-FH10T+ (N term)
pMCSG12 (linearized)
pMCSG35B
pMCSG62 (linearized)
pSpeedET
pMCSG13
pMCSG35B (linearized)
pMCSG63
pVP13
pMCSG13 (linearized)
pMCSG37
pMCSG63 (linearized)
pVP16
pMCSG14
pMCSG37 (linearized)
pMCSG64
pVP33A
pMCSG14 (linearized)
pMCSG38
pMCSG64 (linearized)
pVP33K
pMCSG17
pMCSG38 (linearized)
pMCSG65
pVP56A
pMCSG17 (linearized)
pMCSG38C
pMCSG65 (linearized)
pVP56K
pMCSG18
pMCSG38C (linearized)
pMCSG66
pVP65K
pMCSG18 (linearized)
pMCSG39
pMCSG66 (linearized)
pVP68K
pMCSG19
pMCSG39 (linearized)
pMCSG67
pVP81K
pMCSG19 (linearized)
pMCSG40
pMCSG67 (linearized)
pVP91A
pMCSG19B
pMCSG40 (linearized)
pMCSG68

Plasmid Sets

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pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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