pEU-GST-FV

Flexi® Vector with the barnase gene, for in vitro transcription and translation of a protein with an N-terminal GST-HRV 3C cassette.

Sequence Author: Center for Eukaryotic Structural Genomics

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HomePlasmidsStructural Genomics VectorspEU-GST-FV
BsrGI (5549) NaeI (5356) NgoMIV (5354) SgrAI (5345) AfeI (5263) PfoI (4964) FspI (4153) BglI (4051) BsaI (3992) AhdI (3931) AlwNI (3454) BsgI (438) SwaI (579) BclI * (586) AsiSI - SgfI (840) Bpu10I (1288) BspEI (1511) PasI - PflMI * (1747) PmeI (1966) Eco53kI (1979) SacI (1981) Acc65I (1983) KpnI (1987) XbaI (1998) SalI (2004) AccI (2005) HincII (2006) PstI - SbfI (2014) BlpI (2079) SpeI (2097) PshAI (2127) XcmI (2387) NsiI (2435) PmlI (2474) BsiWI - SnaBI (2481) BssHII (2538) HindIII (2679) pEU-GST-FV 5612 bp
BsrGI  (5549)
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.
NaeI  (5356)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (5354)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
SgrAI  (5345)
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.
AfeI  (5263)
1 site		 A G C G C T T C G C G A
PfoI  (4964)
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.
FspI  (4153)
1 site		 T G C G C A A C G C G T
BglI  (4051)
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.
BsaI  (3992)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (3931)
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  (3454)
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.
BsgI  (438)
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).
SwaI  (579)
1 site		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
BclI  (586)
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.
AsiSI  (840)
1 site		 G C G A T C G C C G C T A G C G
SgfI  (840)
1 site		 G C G A T C G C C G C T A G C G
Bpu10I  (1288)
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.
BspEI  (1511)
1 site		 T C C G G A A G G C C T
PasI  (1747)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
PflMI  (1747)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
PmeI  (1966)
1 site		 G T T T A A A C C A A A T T T G
Eco53kI  (1979)
1 site		 G A G C T C C T C G A G
SacI  (1981)
1 site		 G A G C T C C T C G A G
Acc65I  (1983)
1 site		 G G T A C C C C A T G G
KpnI  (1987)
1 site		 G G T A C C C C A T G G
XbaI  (1998)
1 site		 T C T A G A A G A T C T
SalI  (2004)
1 site		 G T C G A C C A G C T G
AccI  (2005)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HincII  (2006)
1 site		 G T Y R A C C A R Y T G
PstI  (2014)
1 site		 C T G C A G G A C G T C
SbfI  (2014)
1 site		 C C T G C A G G G G A C G T C C
BlpI  (2079)
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.
SpeI  (2097)
1 site		 A C T A G T T G A T C A
PshAI  (2127)
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.
XcmI  (2387)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
NsiI  (2435)
1 site		 A T G C A T T A C G T A
PmlI  (2474)
1 site		 C A C G T G G T G C A C
BsiWI  (2481)
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.
SnaBI  (2481)
1 site		 T A C G T A A T G C A T
BssHII  (2538)
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.
HindIII  (2679)
1 site		 A A G C T T T T C G A A
AmpR
3858 .. 4718  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3858 .. 4649  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3858 .. 4718  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4650 .. 4718  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3858 .. 4718  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
GST
149 .. 805  =  657 bp
219 amino acids  =  25.6 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
149 .. 805  =  657 bp
219 amino acids  =  25.6 kDa
Product: glutathione S-transferase from Schistosoma japonicum
HRV 3C site
812 .. 835  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
HRV 3C site
812 .. 835  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
CmR
1302 .. 1961  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1302 .. 1961  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3099 .. 3687  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3099 .. 3687  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
867 .. 1202  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
867 .. 1202  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
AmpR promoter
4719 .. 4823  =  105 bp
AmpR promoter
4719 .. 4823  =  105 bp
lac UV5 promoter
1218 .. 1248  =  31 bp
3 segments
   Segment 1:  -35  
   1218 .. 1223  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1218 .. 1248  =  31 bp
3 segments
   Segment 2:  
   1224 .. 1241  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1218 .. 1248  =  31 bp
3 segments
   Segment 3:  -10  
   1242 .. 1248  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1218 .. 1248  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
SP6 promoter
1 .. 19  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
1 .. 19  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
ORF:  751 .. 1155  =  405 bp
ORF:  134 amino acids  =  14.7 kDa
ORF:  3988 .. 4254  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  149 .. 853  =  705 bp
ORF:  234 amino acids  =  27.2 kDa
ORF:  5195 .. 5485  =  291 bp
ORF:  96 amino acids  =  10.2 kDa
ORF:  867 .. 1202  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  1302 .. 1961  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3858 .. 4718  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  5060 .. 5422  =  363 bp
ORF:  120 amino acids  =  12.2 kDa
ORF:  589 .. 846  =  258 bp
ORF:  85 amino acids  =  9.4 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

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