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Plasmid Files

pMMB67HE

Broad-host-range bacterial cloning vector with the pUC18/19 MCS and an ampicillin resistance gene. See also pMMB67EH.

 
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pMMB67HE.dna
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PacI (3) MauBI (8564) MluI (8107) ApaI (7904) PspOMI (7900) HpaI (7605) BsmBI (7492) BspQI - SapI (7299) FspAI (6828) BsaAI (6773) EcoNI (6678) PfoI * (6538) SgrAI (4590) HindIII (5) SphI (15) PstI - SbfI (21) SalI (23) XbaI (29) BamHI (35) TspMI - XmaI (40) SmaI (42) Acc65I (44) KpnI (48) Eco53kI (52) SacI (54) EcoRI (56) BssS α I (703) DraI (794) PvuI (1003) AhdI (1372) SexAI * (1496) SfiI (1730) EcoO109I (2282) BstZ17I (3105) PasI (4111) pMMB67HE 8843 bp
PacI  (3)
1 site
T T A A T T A A A A T T A A T T
MauBI  (8564)
1 site
C G C G C G C G G C G C G C G C
MluI  (8107)
1 site
A C G C G T T G C G C A
ApaI  (7904)
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  (7900)
1 site
G G G C C C C C C G G G
HpaI  (7605)
1 site
G T T A A C C A A T T G
BsmBI  (7492)
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.
BspQI  (7299)
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  (7299)
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  (6828)
1 site
R T G C G C A Y Y A C G C G T R
BsaAI  (6773)
1 site
Y A C G T R R T G C A Y
EcoNI  (6678)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PfoI  (6538)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
SgrAI  (4590)
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.
HindIII  (5)
1 site
A A G C T T T T C G A A
SphI  (15)
1 site
G C A T G C C G T A C G
PstI  (21)
1 site
C T G C A G G A C G T C
SbfI  (21)
1 site
C C T G C A G G G G A C G T C C
SalI  (23)
1 site
G T C G A C C A G C T G
XbaI  (29)
1 site
T C T A G A A G A T C T
BamHI  (35)
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.
TspMI  (40)
1 site
C C C G G G G G G C C C
XmaI  (40)
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  (42)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (44)
1 site
G G T A C C C C A T G G
KpnI  (48)
1 site
G G T A C C C C A T G G
Eco53kI  (52)
1 site
G A G C T C C T C G A G
SacI  (54)
1 site
G A G C T C C T C G A G
EcoRI  (56)
1 site
G A A T T C C T T A A G
BssSαI  (703)
1 site
C A C G A G G T G C T C
DraI  (794)
1 site
T T T A A A A A A T T T
PvuI  (1003)
1 site
C G A T C G G C T A G C
AhdI  (1372)
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.
SexAI  (1496)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (1730)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
EcoO109I  (2282)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BstZ17I  (3105)
1 site
G T A T A C C A T A T G
PasI  (4111)
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.
lacI
7380 .. 8462  =  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
7380 .. 8462  =  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).
RSF1010 RepB
3944 .. 4915  =  972 bp
323 amino acids  =  35.9 kDa
Product: replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepB
3944 .. 4915  =  972 bp
323 amino acids  =  35.9 kDa
Product: replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
AmpR
585 .. 1445  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   585 .. 653  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
585 .. 1445  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   654 .. 1445  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
585 .. 1445  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
RSF1010 RepC
6252 .. 7103  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepC
6252 .. 7103  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriV
1882 .. 2276  =  395 bp
replication origin of the broad-host-range plasmid RSF1010; requires the RSF1010 RepA/B/C proteins for replication (Scholz et al., 1989)
RSF1010 oriV
1882 .. 2276  =  395 bp
replication origin of the broad-host-range plasmid RSF1010; requires the RSF1010 RepA/B/C proteins for replication (Scholz et al., 1989)
AmpR promoter
493 .. 584  =  92 bp
AmpR promoter
493 .. 584  =  92 bp
RSF1010 oriT
2618 .. 2705  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriT
2618 .. 2705  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
rrnB T1 terminator
269 .. 355  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
269 .. 355  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
lacIq promoter
8463 .. 8540  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
8463 .. 8540  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
MCS
5 .. 61  =  57 bp
pUC18/19 multiple cloning site
MCS
5 .. 61  =  57 bp
pUC18/19 multiple cloning site
tac promoter
8770 .. 8798  =  29 bp
   Segment 1:  -35  
   8770 .. 8775  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8770 .. 8798  =  29 bp
   Segment 2:  
   8776 .. 8791  =  16 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8770 .. 8798  =  29 bp
   Segment 3:  -10  
   8792 .. 8798  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8770 .. 8798  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
rrnB T2 terminator
447 .. 474  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
447 .. 474  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
lac operator
8806 .. 8822  =  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
8806 .. 8822  =  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).
RSF1010 RepA
5426 .. 6265  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepA
5426 .. 6265  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
ORF:  1543 .. 1875  =  333 bp
ORF:  110 amino acids  =  12.4 kDa
ORF:  2786 .. 4915  =  2130 bp
ORF:  709 amino acids  =  78.0 kDa
ORF:  5426 .. 6265  =  840 bp
ORF:  279 amino acids  =  29.9 kDa
ORF:  585 .. 1445  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2082 .. 2510  =  429 bp
ORF:  142 amino acids  =  15.3 kDa
ORF:  2628 .. 3035  =  408 bp
ORF:  135 amino acids  =  14.3 kDa
ORF:  3534 .. 3947  =  414 bp
ORF:  137 amino acids  =  15.1 kDa
ORF:  4371 .. 5015  =  645 bp
ORF:  214 amino acids  =  22.8 kDa
ORF:  6153 .. 7103  =  951 bp
ORF:  316 amino acids  =  34.4 kDa
ORF:  1464 .. 1742  =  279 bp
ORF:  92 amino acids  =  10.3 kDa
ORF:  6486 .. 6824  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
ORF:  7380 .. 8339  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  110 .. 436  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  1049 .. 1315  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2303 .. 2587  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  2657 .. 3313  =  657 bp
ORF:  218 amino acids  =  22.9 kDa
ORF:  3506 .. 3844  =  339 bp
ORF:  112 amino acids  =  12.0 kDa
ORF:  4061 .. 4654  =  594 bp
ORF:  197 amino acids  =  20.5 kDa
ORF:  5423 .. 6190  =  768 bp
ORF:  255 amino acids  =  27.0 kDa
ORF:  6593 .. 6934  =  342 bp
ORF:  113 amino acids  =  12.6 kDa
ORF:  1432 .. 1749  =  318 bp
ORF:  105 amino acids  =  12.2 kDa
ORF:  2953 .. 3183  =  231 bp
ORF:  76 amino acids  =  8.4 kDa
ORF:  3850 .. 4203  =  354 bp
ORF:  117 amino acids  =  12.6 kDa
ORF:  5677 .. 6042  =  366 bp
ORF:  121 amino acids  =  13.3 kDa
ORF:  7360 .. 7623  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  8212 .. 8529  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
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