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Plasmid Files

pAB-6xHis™

Baculovirus transfer plasmid for high-level expression of proteins with a thrombin-cleavable N-terminal 6xHis tag.

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pAB-6xHis.dna
Map and Sequence File:    Download    Open   
Sequence Author:  AB Vector
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NdeI (9484) ScaI (8793) BpmI (8383) AlwNI (7836) PspFI (7728) BseYI (7724) BspQI - SapI (7304) FspAI (6387) SgrDI (6234) SgrAI (5965) SnaBI (4877) PasI (486) PacI (583) XcmI (746) BtgI (868) SacII (871) PspOMI (1395) ApaI - BanII (1399) StyI (1501) PaeR7I - PspXI - XhoI (1901) SphI (2135) BbvCI - Bpu10I (3405) NgoMIV (3770) NaeI (3772) EcoRV (4000) BamHI (4129) SpeI (4173) TspMI - XmaI (4192) SmaI (4194) XbaI (4203) EcoRI (4207) EagI - NotI (4218) BglII - PstI (4228) KflI - PpuMI (4246) pAB-6xHis™ 9698 bp
NdeI  (9484)
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.
ScaI  (8793)
1 site
A G T A C T T C A T G A
BpmI  (8383)
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.
AlwNI  (7836)
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  (7728)
1 site
C C C A G C G G G T C G
BseYI  (7724)
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.
BspQI  (7304)
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  (7304)
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.
FspAI  (6387)
1 site
R T G C G C A Y Y A C G C G T R
SgrDI  (6234)
1 site
C G T C G A C G G C A G C T G C
SgrAI  (5965)
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.
SnaBI  (4877)
1 site
T A C G T A A T G C A T
PasI  (486)
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.
PacI  (583)
1 site
T T A A T T A A A A T T A A T T
XcmI  (746)
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.
BtgI  (868)
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  (871)
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.
PspOMI  (1395)
1 site
G G G C C C C C C G G G
ApaI  (1399)
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  (1399)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
StyI  (1501)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PaeR7I  (1901)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1901)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1901)
1 site
C T C G A G G A G C T C
SphI  (2135)
1 site
G C A T G C C G T A C G
BbvCI  (3405)
1 site
C C T C A G C G G A G T C G
Bpu10I  (3405)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
NgoMIV  (3770)
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.
NaeI  (3772)
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.
EcoRV  (4000)
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.
BamHI  (4129)
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.
SpeI  (4173)
1 site
A C T A G T T G A T C A
TspMI  (4192)
1 site
C C C G G G G G G C C C
XmaI  (4192)
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  (4194)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
XbaI  (4203)
1 site
T C T A G A A G A T C T
EcoRI  (4207)
1 site
G A A T T C C T T A A G
EagI  (4218)
1 site
C G G C C G G C C G G C
NotI  (4218)
1 site
G C G G C C G C C G C C G G C G
BglII  (4228)
1 site
A G A T C T T C T A G A
PstI  (4228)
1 site
C T G C A G G A C G T C
KflI  (4246)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (4246)
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.
AmpR
8240 .. 9100  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   8240 .. 9031  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8240 .. 9100  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   9032 .. 9100  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8240 .. 9100  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
baculovirus recombination region (lef2/ORF603)
3170 .. 3997  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
3170 .. 3997  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (ORF1629)
4713 .. 5418  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4713 .. 5418  =  706 bp
contains part of ORF1629
ori
7481 .. 8069  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7481 .. 8069  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
9101 .. 9205  =  105 bp
AmpR promoter
9101 .. 9205  =  105 bp
polyhedrin promoter
4001 .. 4092  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4001 .. 4092  =  92 bp
promoter for the baculovirus polyhedrin gene
MCS
4192 .. 4251  =  60 bp
multiple cloning site
MCS
4192 .. 4251  =  60 bp
multiple cloning site
6xHis
4147 .. 4164  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4147 .. 4164  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
thrombin site
4174 .. 4191  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
4174 .. 4191  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
ATG
4138 .. 4140  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4138 .. 4140  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF603
3332 .. 3937  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
3332 .. 3937  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF1629
4830 .. 5418  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4830 .. 5418  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
lef2
3170 .. 3294  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus