pIB V5-His-DEST

Gateway® destination vector for stable constitutive expression of C-terminally V5-6xHis-tagged proteins in insect cells.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspIB V5-His-DEST
AhdI (4870) BglI (4752) XmnI (4270) EarI (4071) BglII (4003) DraIII (3970) BlpI (3960) NruI * (3796) PpuMI (3724) MreI - SgrAI (3658) BbsI (3650) SpeI (3415) BciVI (2826) PciI (2623) BspDI * - ClaI * (2560) BspHI (0) FseI (293) BsiWI (317) RsrII (335) KasI (474) NarI (475) SfoI (476) PluTI (478) HindIII (557) EcoRV (565) AfeI (586) NotI (728) BspEI (1044) BsmBI - Esp3I (1272) PasI - PflMI * (1280) SspI (1360) BssHII (1542) BbvCI (1773) SrfI (1919) BstXI (2036) BfuAI - BspMI (2163) SalI (2176) PshAI (2315) XbaI (2319) SacII (2332) AgeI (2386) 6xHis PsiI (2493) pIB/V5-His-DEST 5038 bp
AhdI  (4870)
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.
BglI  (4752)
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.
XmnI  (4270)
1 site		 G A A N N N N T T C C T T N N N N A A G
EarI  (4071)
1 site		 C T C T T C N G A G A A G N N N N
Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
BglII  (4003)
1 site		 A G A T C T T C T A G A
DraIII  (3970)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
BlpI  (3960)
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.
NruI  (3796)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
PpuMI  (3724)
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.
MreI  (3658)
1 site		 C G C C G G C G G C G G C C G C
SgrAI  (3658)
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.
BbsI  (3650)
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.
SpeI  (3415)
1 site		 A C T A G T T G A T C A
BciVI  (2826)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
PciI  (2623)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspDI  (2560)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2560)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
BspHI  (0)
1 site		 T C A T G A A G T A C T
FseI  (293)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
BsiWI  (317)
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.
RsrII  (335)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
KasI  (474)
1 site		 G G C G C C C C G C G G
NarI  (475)
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  (476)
1 site		 G G C G C C C C G C G G
PluTI  (478)
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.
HindIII  (557)
1 site		 A A G C T T T T C G A A
EcoRV  (565)
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.
AfeI  (586)
1 site		 A G C G C T T C G C G A
NotI  (728)
1 site		 G C G G C C G C C G C C G G C G
BspEI  (1044)
1 site		 T C C G G A A G G C C T
BsmBI  (1272)
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  (1272)
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.
PasI  (1280)
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  (1280)
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.
SspI  (1360)
1 site		 A A T A T T T T A T A A
BssHII  (1542)
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.
BbvCI  (1773)
1 site		 C C T C A G C G G A G T C G
SrfI  (1919)
1 site		 G C C C G G G C C G G G C C C G
BstXI  (2036)
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.
BfuAI  (2163)
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  (2163)
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.
SalI  (2176)
1 site		 G T C G A C C A G C T G
PshAI  (2315)
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.
XbaI  (2319)
1 site		 T C T A G A A G A T C T
SacII  (2332)
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.
AgeI  (2386)
1 site		 A C C G G T T G G C C A
PsiI  (2493)
1 site		 T T A T A A A A T A T T
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4083 .. 4151  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4152 .. 4943  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
835 .. 1494  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
835 .. 1494  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2684 .. 3272  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2684 .. 3272  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
OpIE-2 promoter
1 .. 548  =  548 bp
strong constitutive baculovirus promoter for insect cell expression
OpIE-2 promoter
1 .. 548  =  548 bp
strong constitutive baculovirus promoter for insect cell expression
BSD
3565 .. 3963  =  399 bp
132 amino acids  =  13.6 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
3565 .. 3963  =  399 bp
132 amino acids  =  13.6 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
ccdB
1836 .. 2141  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
ccdB
1836 .. 2141  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
OpIE-2 poly(A) signal
2427 .. 2556  =  130 bp
baculovirus polyadenylation signal
OpIE-2 poly(A) signal
2427 .. 2556  =  130 bp
baculovirus polyadenylation signal
attR1
602 .. 726  =  125 bp
recombination site for the Gateway® LR reaction
attR1
602 .. 726  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2182 .. 2306  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2182 .. 2306  =  125 bp
recombination site for the Gateway® LR reaction
gp64 promoter
3364 .. 3463  =  100 bp
baculovirus promoter for constitutive expression in insect cells
gp64 promoter
3364 .. 3463  =  100 bp
baculovirus promoter for constitutive expression in insect cells
V5 tag
2341 .. 2382  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2341 .. 2382  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
2392 .. 2409  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2392 .. 2409  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
EM7 promoter
3499 .. 3546  =  48 bp
synthetic bacterial promoter
EM7 promoter
3499 .. 3546  =  48 bp
synthetic bacterial promoter
lac UV5 promoter
751 .. 781  =  31 bp
3 segments
   Segment 1:  -35  
   751 .. 756  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
751 .. 781  =  31 bp
3 segments
   Segment 2:  
   757 .. 774  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
751 .. 781  =  31 bp
3 segments
   Segment 3:  -10  
   775 .. 781  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
751 .. 781  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
ORF:  835 .. 1494  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3565 .. 3963  =  399 bp
ORF:  132 amino acids  =  13.6 kDa
ORF:  15 .. 245  =  231 bp
ORF:  76 amino acids  =  8.9 kDa
ORF:  1836 .. 2141  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4083 .. 4943  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4547 .. 4813  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3511 .. 3867  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
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Download pIB V5-His-DEST.dna file

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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
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Plant Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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