pVP16

Bacterial Gateway® destination vector for appending an N-terminal 8xHis-MBP cassette to an expressed protein.

Sequence Author: Center for Eukaryotic Structural Genomics

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HomePlasmidsStructural Genomics VectorspVP16
PaeR7I - XhoI (2) XmnI (7238) BglI (6759) AhdI (6639) PciI (5746) BspQI - SapI (5630) NdeI (5569) PflFI - Tth111I (5493) FspAI (5326) PpuMI * (5309) BstEII (4674) ApaI (4653) PspOMI (4649) EcoRV (4410) HpaI (4354) PluTI (4221) SfoI (4219) NarI * (4218) KasI (4217) T5 promoter MfeI (60) lac operator RBS BseRI (116) ATG BsiWI (436) AfeI (1056) Eco53kI (1254) SacI (1256) EagI - NotI (1442) BbvCI (2487) TspMI - XmaI (2631) SmaI - SrfI (2633) PstI (2888) AscI (3025) SgrDI (3040) Acc65I (3046) KpnI (3050) BspDI - ClaI (3053) BstBI (3064) HindIII (3067) NheI (3187) BmtI (3191) pVP16 7631 bp
PaeR7I  (2)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2)
1 site		 C T C G A G G A G C T C
XmnI  (7238)
1 site		 G A A N N N N T T C C T T N N N N A A G
BglI  (6759)
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  (6639)
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.
PciI  (5746)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (5630)
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  (5630)
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.
NdeI  (5569)
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.
PflFI  (5493)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (5493)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
FspAI  (5326)
1 site		 R T G C G C A Y Y A C G C G T R
PpuMI  (5309)
1 site		 R G G W C C Y Y C C W G G R
* Blocked by Dcm methylation.
Sticky ends from different PpuMI sites may not be compatible.
BstEII  (4674)
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.
ApaI  (4653)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (4649)
1 site		 G G G C C C C C C G G G
EcoRV  (4410)
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  (4354)
1 site		 G T T A A C C A A T T G
PluTI  (4221)
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.
SfoI  (4219)
1 site		 G G C G C C C C G C G G
NarI  (4218)
1 site		 G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (4217)
1 site		 G G C G C C C C G C G G
MfeI  (60)
1 site		 C A A T T G G T T A A C
BseRI  (116)
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.
BsiWI  (436)
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.
AfeI  (1056)
1 site		 A G C G C T T C G C G A
Eco53kI  (1254)
1 site		 G A G C T C C T C G A G
SacI  (1256)
1 site		 G A G C T C C T C G A G
EagI  (1442)
1 site		 C G G C C G G C C G G C
NotI  (1442)
1 site		 G C G G C C G C C G C C G G C G
BbvCI  (2487)
1 site		 C C T C A G C G G A G T C G
TspMI  (2631)
1 site		 C C C G G G G G G C C C
XmaI  (2631)
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.
SmaI  (2633)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
SrfI  (2633)
1 site		 G C C C G G G C C G G G C C C G
PstI  (2888)
1 site		 C T G C A G G A C G T C
AscI  (3025)
1 site		 G G C G C G C C C C G C G C G G
SgrDI  (3040)
1 site		 C G T C G A C G G C A G C T G C
Acc65I  (3046)
1 site		 G G T A C C C C A T G G
KpnI  (3050)
1 site		 G G T A C C C C A T G G
BspDI  (3053)
1 site		 A T C G A T T A G C T A
ClaI  (3053)
1 site		 A T C G A T T A G C T A
BstBI  (3064)
1 site		 T T C G A A A A G C T T
HindIII  (3067)
1 site		 A A G C T T T T C G A A
NheI  (3187)
1 site		 G C T A G C C G A T C G
BmtI  (3191)
1 site		 G C T A G C C G A T C G
ATG
116 .. 118  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
116 .. 118  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
8xHis
122 .. 145  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
122 .. 145  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
MBP
149 .. 1246  =  1098 bp
366 amino acids  =  40.2 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
149 .. 1246  =  1098 bp
366 amino acids  =  40.2 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.
lacI
4129 .. 5211  =  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
4129 .. 5211  =  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
6566 .. 7426  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6566 .. 7357  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6566 .. 7426  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7358 .. 7426  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6566 .. 7426  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
1549 .. 2208  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1549 .. 2208  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3226 .. 3885  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3226 .. 3885  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5807 .. 6395  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5807 .. 6395  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ccdB
2550 .. 2855  =  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
2550 .. 2855  =  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.
attR1
1316 .. 1440  =  125 bp
recombination site for the Gateway® LR reaction
attR1
1316 .. 1440  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2896 .. 3020  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2896 .. 3020  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
7427 .. 7531  =  105 bp
AmpR promoter
7427 .. 7531  =  105 bp
lambda t0 terminator
3088 .. 3182  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
3088 .. 3182  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
3950 .. 4036  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3950 .. 4036  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
lacIq promoter
5212 .. 5289  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
5212 .. 5289  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
T5 promoter
11 .. 55  =  45 bp
4 segments
   Segment 1:  
   11 .. 25  =  15 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
11 .. 55  =  45 bp
4 segments
   Segment 2:  -35  
   26 .. 31  =  6 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
11 .. 55  =  45 bp
4 segments
   Segment 3:  
   32 .. 48  =  17 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
11 .. 55  =  45 bp
4 segments
   Segment 4:  -10  
   49 .. 55  =  7 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
11 .. 55  =  45 bp
4 segments
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
lac UV5 promoter
1465 .. 1495  =  31 bp
3 segments
   Segment 1:  -35  
   1465 .. 1470  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1465 .. 1495  =  31 bp
3 segments
   Segment 2:  
   1471 .. 1488  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1465 .. 1495  =  31 bp
3 segments
   Segment 3:  -10  
   1489 .. 1495  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1465 .. 1495  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac operator
63 .. 79  =  17 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
63 .. 79  =  17 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).
RBS
102 .. 107  =  6 bp
ribosome binding site
RBS
102 .. 107  =  6 bp
ribosome binding site
lac operator
31 .. 47  =  17 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
31 .. 47  =  17 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).
ORF:  1549 .. 2208  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3226 .. 3885  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  116 .. 1351  =  1236 bp
ORF:  411 amino acids  =  45.4 kDa
ORF:  2550 .. 2855  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4095 .. 4346  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  6696 .. 6962  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4109 .. 4372  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4961 .. 5278  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  6566 .. 7426  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4129 .. 5088  =  960 bp
ORF:  319 amino acids  =  34.1 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

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
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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