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Plasmid Files

pMAL-p5E

Bacterial vector for inducible periplasmic expression of maltose-binding protein (MBP) fusions with an enterokinase cleavage site.

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pMAL-p5E Sequence and MappMAL-p5E.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 FspAI (5720) Bpu10I (5592) PflFI - Tth111I (4955) BspQI - SapI (4874) PciI (4757) Bsu36I (4068) BglI (3834) ScaI (3471) TatI (3469) DraI (3374) BspHI (3112) RsrII (3068) rrnB T2 terminator MscI * (5730) PflMI (13) MluI (431) BstEII (612) PspOMI (638) ApaI (642) HpaI (937) KasI (1070) NarI * (1071) SfoI (1072) PluTI (1074) lac operator MfeI (1502) BsiWI (1893) PsiI (1955) BglII (1962) BmgBI (2145) BsmI (2223) BlpI (2389) Eco53kI (2711) SacI (2713) AvaI - BsoBI (2746) BmeT110I (2747) Acc65I (2766) KpnI (2770) NdeI (2774) NcoI - StyI (2780) EagI - NotI (2787) EcoRV (2796) SalI - SgrDI (2800) AccI (2801) BamHI (2806) EcoRI (2812) PstI - SbfI (2823) HindIII (2835) pMAL-p5E 5755 bp
FspAI  (5720)
1 site
R T G C G C A Y Y A C G C G T R
Bpu10I  (5592)
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.
PflFI  (4955)
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  (4955)
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.
BspQI  (4874)
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  (4874)
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  (4757)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
Bsu36I  (4068)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BglI  (3834)
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.
ScaI  (3471)
1 site
A G T A C T T C A T G A
TatI  (3469)
1 site
W G T A C W W C A T G W
DraI  (3374)
1 site
T T T A A A A A A T T T
BspHI  (3112)
1 site
T C A T G A A G T A C T
RsrII  (3068)
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.
MscI  (5730)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
PflMI  (13)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
MluI  (431)
1 site
A C G C G T T G C G C A
BstEII  (612)
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.
PspOMI  (638)
1 site
G G G C C C C C C G G G
ApaI  (642)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
HpaI  (937)
1 site
G T T A A C C A A T T G
KasI  (1070)
1 site
G G C G C C C C G C G G
NarI  (1071)
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.
SfoI  (1072)
1 site
G G C G C C C C G C G G
PluTI  (1074)
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.
MfeI  (1502)
1 site
C A A T T G G T T A A C
BsiWI  (1893)
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.
PsiI  (1955)
1 site
T T A T A A A A T A T T
BglII  (1962)
1 site
A G A T C T T C T A G A
BmgBI  (2145)
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.
BsmI  (2223)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BlpI  (2389)
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.
Eco53kI  (2711)
1 site
G A G C T C C T C G A G
SacI  (2713)
1 site
G A G C T C C T C G A G
AvaI  (2746)
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  (2746)
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.
BmeT110I  (2747)
1 site
C Y C G R G G R G C Y C
Acc65I  (2766)
1 site
G G T A C C C C A T G G
KpnI  (2770)
1 site
G G T A C C C C A T G G
NdeI  (2774)
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.
NcoI  (2780)
1 site
C C A T G G G G T A C C
StyI  (2780)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
EagI  (2787)
1 site
C G G C C G G C C G G C
NotI  (2787)
1 site
G C G G C C G C C G C C G G C G
EcoRV  (2796)
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.
SalI  (2800)
1 site
G T C G A C C A G C T G
SgrDI  (2800)
1 site
C G T C G A C G G C A G C T G C
AccI  (2801)
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.
BamHI  (2806)
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.
EcoRI  (2812)
1 site
G A A T T C C T T A A G
PstI  (2823)
1 site
C T G C A G G A C G T C
SbfI  (2823)
1 site
C C T G C A G G G G A C G T C C
HindIII  (2835)
1 site
A A G C T T T T C G A A
MBP
1528 .. 2703  =  1176 bp
392 amino acids  =  42.9 kDa
   Segment 1:  signal sequence  
   1528 .. 1605  =  78 bp
   26 amino acids  =  2.7 kDa
Product: maltose binding protein from E. coli
This gene encodes a signal sequence that can direct
MBP to the bacterial periplasm.
MBP
1528 .. 2703  =  1176 bp
392 amino acids  =  42.9 kDa
   Segment 2:  
   1606 .. 2703  =  1098 bp
   366 amino acids  =  40.2 kDa
Product: maltose binding protein from E. coli
This gene encodes a signal sequence that can direct
MBP to the bacterial periplasm.
MBP
1528 .. 2703  =  1176 bp
392 amino acids  =  42.9 kDa
2 segments
Product: maltose binding protein from E. coli
This gene encodes a signal sequence that can direct
MBP to the bacterial periplasm.
lacI
81 .. 1163  =  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
81 .. 1163  =  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
3165 .. 4025  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3165 .. 3233  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3165 .. 4025  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3234 .. 4025  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3165 .. 4025  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
4113 .. 4701  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4113 .. 4701  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
rop
5071 .. 5262  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
5071 .. 5262  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
AmpR promoter
3074 .. 3164  =  91 bp
AmpR promoter
3074 .. 3164  =  91 bp
rrnB T1 terminator
2841 .. 2927  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
2841 .. 2927  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
lacIq promoter
3 .. 80  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
lacIq promoter
3 .. 80  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
MCS
2766 .. 2840  =  75 bp
multiple cloning site
MCS
2766 .. 2840  =  75 bp
multiple cloning site
tac promoter
1406 .. 1434  =  29 bp
   Segment 1:  -35  
   1406 .. 1411  =  6 bp
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
tac promoter
1406 .. 1434  =  29 bp
   Segment 2:  
   1412 .. 1427  =  16 bp
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
tac promoter
1406 .. 1434  =  29 bp
   Segment 3:  -10  
   1428 .. 1434  =  7 bp
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
tac promoter
1406 .. 1434  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
rrnB T2 terminator
3019 .. 3046  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
3019 .. 3046  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
lac operator
1442 .. 1458  =  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
1442 .. 1458  =  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).
enterokinase site
2752 .. 2766  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
2752 .. 2766  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
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