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pAc5.1/V5-His A

Vector for constitutive expression of C-terminally V5-6xHis-tagged proteins in insect cells. For other reading frames, use pAc5.1/V5‑His B or pAc5.1/V5‑His C.

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pAc5.1 V5-His A Sequence and MappAc5.1 V5-His A.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 ScaI (4869) NmeAIII (4537) AhdI (4389) DrdI (3604) BfuAI - BspMI (3446) PshAI (3108) SalI (3043) HpaI (2935) AanI (2915) BamHI (2800) SacI (2779) Eco53kI (2777) StuI (2771) PmeI (2735) AgeI (2705) MluI (2699) BglII (132) PflMI * (383) AsiSI (391) TspMI - XmaI (425) SmaI (427) BbvCI (502) AanI (692) BsiWI (1096) SphI (1105) KasI (1388) NarI (1389) SfoI (1390) PluTI (1392) NsiI (1406) BspDI - ClaI (1708) AflII (1934) NdeI (2120) PpuMI (2257) BtgI - NcoI - StyI (2261) BlpI (2439) AanI (2452) Acc65I (2579) KpnI (2583) BstXI (2599) EcoRI (2603) EcoRV (2615) BstXI (2625) NotI (2630) PaeR7I - PspXI - XhoI (2636) XbaI (2642) PspOMI (2648) ApaI (2652) BstBI (2655) V5 tag pAc5.1/V5-His A 5375 bp
ScaI  (4869)
1 site
A G T A C T T C A T G A
NmeAIII  (4537)
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).
AhdI  (4389)
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.
DrdI  (3604)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BfuAI  (3446)
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  (3446)
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.
PshAI  (3108)
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.
SalI  (3043)
1 site
G T C G A C C A G C T G
HpaI  (2935)
1 site
G T T A A C C A A T T G
AanI  (2915)
3 sites
T T A T A A A A T A T T
BamHI  (2800)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
SacI  (2779)
1 site
G A G C T C C T C G A G
Eco53kI  (2777)
1 site
G A G C T C C T C G A G
StuI  (2771)
1 site
A G G C C T T C C G G A
PmeI  (2735)
1 site
G T T T A A A C C A A A T T T G
AgeI  (2705)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
MluI  (2699)
1 site
A C G C G T T G C G C A
BglII  (132)
1 site
A G A T C T T C T A G A
PflMI  (383)
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.
AsiSI  (391)
1 site
G C G A T C G C C G C T A G C G
TspMI  (425)
1 site
C C C G G G G G G C C C
XmaI  (425)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (427)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BbvCI  (502)
1 site
C C T C A G C G G A G T C G
AanI  (692)
3 sites
T T A T A A A A T A T T
BsiWI  (1096)
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.
SphI  (1105)
1 site
G C A T G C C G T A C G
KasI  (1388)
1 site
G G C G C C C C G C G G
NarI  (1389)
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  (1390)
1 site
G G C G C C C C G C G G
PluTI  (1392)
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.
NsiI  (1406)
1 site
A T G C A T T A C G T A
BspDI  (1708)
1 site
A T C G A T T A G C T A
ClaI  (1708)
1 site
A T C G A T T A G C T A
AflII  (1934)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
NdeI  (2120)
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.
PpuMI  (2257)
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.
BtgI  (2261)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (2261)
1 site
C C A T G G G G T A C C
StyI  (2261)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BlpI  (2439)
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.
AanI  (2452)
3 sites
T T A T A A A A T A T T
Acc65I  (2579)
1 site
G G T A C C C C A T G G
KpnI  (2583)
1 site
G G T A C C C C A T G G
BstXI  (2599)
2 sites
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.
EcoRI  (2603)
1 site
G A A T T C C T T A A G
EcoRV  (2615)
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.
BstXI  (2625)
2 sites
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.
NotI  (2630)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (2636)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2636)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2636)
1 site
C T C G A G G A G C T C
XbaI  (2642)
1 site
T C T A G A A G A T C T
PspOMI  (2648)
1 site
G G G C C C C C C G G G
ApaI  (2652)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BstBI  (2655)
1 site
T T C G A A A A G C T T
Ac5 promoter
56 .. 2568  =  2513 bp
Drosophila melanogaster actin 5C promoter
Ac5 promoter
56 .. 2568  =  2513 bp
Drosophila melanogaster actin 5C promoter
AmpR
4316 .. 5176  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   4316 .. 5107  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4316 .. 5176  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5108 .. 5176  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4316 .. 5176  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3557 .. 4145  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3557 .. 4145  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 poly(A) signal
2801 .. 2935  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2801 .. 2935  =  135 bp
SV40 polyadenylation signal
AmpR promoter
5177 .. 5281  =  105 bp
AmpR promoter
5177 .. 5281  =  105 bp
MCS
2579 .. 2659  =  81 bp
multiple cloning site
MCS
2579 .. 2659  =  81 bp
multiple cloning site
V5 tag
2660 .. 2701  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2660 .. 2701  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
2711 .. 2728  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2711 .. 2728  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
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