pMT V5-His B

Vector for inducible high-level expression of C-terminally V5-6xHis-tagged proteins in Drosophila cells. For other reading frames, use pMT/V5-His A or pMT/V5-His C.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsInsect Cell VectorspMT V5-His B
AatII (3478) ZraI (3476) SspI (3360) ScaI (3036) NmeAIII (2704) BpmI (2626) AhdI (2556) AlwNI (2079) PspFI (1971) BseYI (1967) PfoI (46) BstAPI (185) KasI (235) NarI (236) SfoI (237) PluTI (239) BsgI (640) BmgBI (662) MscI (771) Acc65I (849) KpnI (853) SpeI (856) EcoRI (873) EcoRV (885) NotI (900) AvaI - BsoBI - PaeR7I - PspXI - XhoI (906) PspOMI (918) ApaI - BanII - BtgI (922) SacII (925) BstBI (929) BseRI (946) MluI (973) AgeI (979) PmeI (1009) PsiI (1158) HpaI (1178) MfeI (1187) SalI (1286) AccI (1287) SbfI (1296) BfuAI - BspMI (1299) BspQI - SapI (1547) PciI (1663) pMT/V5-His B 3542 bp
AatII  (3478)
1 site		 G A C G T C C T G C A G
ZraI  (3476)
1 site		 G A C G T C C T G C A G
SspI  (3360)
1 site		 A A T A T T T T A T A A
ScaI  (3036)
1 site		 A G T A C T T C A T G A
NmeAIII  (2704)
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).
BpmI  (2626)
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.
AhdI  (2556)
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  (2079)
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.
PspFI  (1971)
1 site		 C C C A G C G G G T C G
BseYI  (1967)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PfoI  (46)
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.
BstAPI  (185)
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.
KasI  (235)
1 site		 G G C G C C C C G C G G
NarI  (236)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (237)
1 site		 G G C G C C C C G C G G
PluTI  (239)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
BsgI  (640)
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).
BmgBI  (662)
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.
MscI  (771)
1 site		 T G G C C A A C C G G T
Acc65I  (849)
1 site		 G G T A C C C C A T G G
KpnI  (853)
1 site		 G G T A C C C C A T G G
SpeI  (856)
1 site		 A C T A G T T G A T C A
EcoRI  (873)
1 site		 G A A T T C C T T A A G
EcoRV  (885)
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.
NotI  (900)
1 site		 G C G G C C G C C G C C G G C G
AvaI  (906)
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  (906)
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  (906)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (906)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (906)
1 site		 C T C G A G G A G C T C
PspOMI  (918)
1 site		 G G G C C C C C C G G G
ApaI  (922)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BanII  (922)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
BtgI  (922)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
SacII  (925)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
BstBI  (929)
1 site		 T T C G A A A A G C T T
BseRI  (946)
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.
MluI  (973)
1 site		 A C G C G T T G C G C A
AgeI  (979)
1 site		 A C C G G T T G G C C A
PmeI  (1009)
1 site		 G T T T A A A C C A A A T T T G
PsiI  (1158)
1 site		 T T A T A A A A T A T T
HpaI  (1178)
1 site		 G T T A A C C A A T T G
MfeI  (1187)
1 site		 C A A T T G G T T A A C
SalI  (1286)
1 site		 G T C G A C C A G C T G
AccI  (1287)
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.
SbfI  (1296)
1 site		 C C T G C A G G G G A C G T C C
BfuAI  (1299)
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  (1299)
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.
BspQI  (1547)
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  (1547)
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.
PciI  (1663)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AmpR
2483 .. 3343  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2483 .. 3274  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2483 .. 3343  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3275 .. 3343  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2483 .. 3343  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1724 .. 2312  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1724 .. 2312  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
MT promoter
407 .. 833  =  427 bp
Drosophila metallothionein promoter
MT promoter
407 .. 833  =  427 bp
Drosophila metallothionein promoter
SV40 poly(A) signal
1044 .. 1178  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1044 .. 1178  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3344 .. 3448  =  105 bp
AmpR promoter
3344 .. 3448  =  105 bp
MCS
849 .. 933  =  85 bp
multiple cloning site
MCS
849 .. 933  =  85 bp
multiple cloning site
V5 tag
934 .. 975  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
934 .. 975  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
985 .. 1002  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
985 .. 1002  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ORF:  563 .. 811  =  249 bp
ORF:  82 amino acids  =  9.2 kDa
ORF:  186 .. 446  =  261 bp
ORF:  86 amino acids  =  9.9 kDa
ORF:  2613 .. 2879  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  405 .. 782  =  378 bp
ORF:  125 amino acids  =  13.3 kDa
ORF:  2483 .. 3343  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  466 .. 816  =  351 bp
ORF:  116 amino acids  =  13.3 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

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