pMMB67EH

Broad-host-range bacterial cloning vector with the pUC18/19 MCS and an ampicillin resistance gene. See also pMMB67HE.
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PstI - SbfI (8821) SalI (8811) MCS XbaI (8805) BamHI (8799) SmaI (8796) KpnI - TspMI - XmaI (8794) Acc65I (8790) SacI (8788) Eco53kI (8786) EcoRI (8778) lac operator tac promoter MauBI (8503) lacIq promoter MluI (8046) ApaI (7843) PspOMI (7839) HpaI (7544) BsmBI - Esp3I (7431) BspQI - SapI (7238) FspAI (6767) BsaAI (6712) EcoNI (6617) PfoI * (6477) SgrAI (4529) SphI (8827) HindIII (0) rrnB T1 terminator rrnB T2 terminator BssSI - BssSαI (642) DraI (733) PvuI (942) AhdI (1311) CsiI - SexAI * (1435) SfiI (1669) EcoO109I (2221) BstZ17I (3044) PasI (4050) pMMB67EH 8829 bp
PstI  (8821)
1 site
C T G C A G G A C G T C
SbfI  (8821)
1 site
C C T G C A G G G G A C G T C C
SalI  (8811)
1 site
G T C G A C C A G C T G
XbaI  (8805)
1 site
T C T A G A A G A T C T
BamHI  (8799)
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.
SmaI  (8796)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KpnI  (8794)
1 site
G G T A C C C C A T G G
TspMI  (8794)
1 site
C C C G G G G G G C C C
XmaI  (8794)
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.
Acc65I  (8790)
1 site
G G T A C C C C A T G G
SacI  (8788)
1 site
G A G C T C C T C G A G
Eco53kI  (8786)
1 site
G A G C T C C T C G A G
EcoRI  (8778)
1 site
G A A T T C C T T A A G
MauBI  (8503)
1 site
C G C G C G C G G C G C G C G C
MluI  (8046)
1 site
A C G C G T T G C G C A
ApaI  (7843)
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  (7839)
1 site
G G G C C C C C C G G G
HpaI  (7544)
1 site
G T T A A C C A A T T G
BsmBI  (7431)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (7431)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BspQI  (7238)
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  (7238)
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  (6767)
1 site
R T G C G C A Y Y A C G C G T R
BsaAI  (6712)
1 site
Y A C G T R R T G C A Y
EcoNI  (6617)
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  (6477)
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  (4529)
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.
SphI  (8827)
1 site
G C A T G C C G T A C G
HindIII  (0)
1 site
A A G C T T T T C G A A
BssSI  (642)
1 site
C A C G A G G T G C T C
BssSαI  (642)
1 site
C A C G A G G T G C T C
DraI  (733)
1 site
T T T A A A A A A T T T
PvuI  (942)
1 site
C G A T C G G C T A G C
AhdI  (1311)
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.
CsiI  (1435)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (1435)
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  (1669)
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  (2221)
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  (3044)
1 site
G T A T A C C A T A T G
PasI  (4050)
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
7319 .. 8401  =  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
7319 .. 8401  =  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
3883 .. 4854  =  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
3883 .. 4854  =  972 bp
323 amino acids  =  35.9 kDa
Product: replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
AmpR
524 .. 1384  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   524 .. 592  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
524 .. 1384  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   593 .. 1384  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
524 .. 1384  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
RSF1010 RepC
6191 .. 7042  =  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
6191 .. 7042  =  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
1821 .. 2215  =  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
1821 .. 2215  =  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
432 .. 523  =  92 bp
AmpR promoter
432 .. 523  =  92 bp
RSF1010 oriT
2557 .. 2644  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriT
2557 .. 2644  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
rrnB T1 terminator
208 .. 294  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
208 .. 294  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
lacIq promoter
8402 .. 8479  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
8402 .. 8479  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
MCS
8778 .. 5  =  57 bp
pUC18/19 multiple cloning site
MCS
8778 .. 5  =  57 bp
pUC18/19 multiple cloning site
tac promoter
8709 .. 8737  =  29 bp
3 segments
   Segment 1:  -35  
   8709 .. 8714  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8709 .. 8737  =  29 bp
3 segments
   Segment 2:  
   8715 .. 8730  =  16 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8709 .. 8737  =  29 bp
3 segments
   Segment 3:  -10  
   8731 .. 8737  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8709 .. 8737  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
rrnB T2 terminator
386 .. 413  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
386 .. 413  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
lac operator
8745 .. 8761  =  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
8745 .. 8761  =  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
5365 .. 6204  =  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
5365 .. 6204  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
ORF:  2725 .. 4854  =  2130 bp
ORF:  709 amino acids  =  78.0 kDa
ORF:  5365 .. 6204  =  840 bp
ORF:  279 amino acids  =  29.9 kDa
ORF:  524 .. 1384  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2021 .. 2449  =  429 bp
ORF:  142 amino acids  =  15.3 kDa
ORF:  2567 .. 2974  =  408 bp
ORF:  135 amino acids  =  14.3 kDa
ORF:  3473 .. 3886  =  414 bp
ORF:  137 amino acids  =  15.1 kDa
ORF:  4310 .. 4954  =  645 bp
ORF:  214 amino acids  =  22.8 kDa
ORF:  6092 .. 7042  =  951 bp
ORF:  316 amino acids  =  34.4 kDa
ORF:  1482 .. 1814  =  333 bp
ORF:  110 amino acids  =  12.4 kDa
ORF:  49 .. 375  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  988 .. 1254  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2242 .. 2526  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  2596 .. 3252  =  657 bp
ORF:  218 amino acids  =  22.9 kDa
ORF:  3445 .. 3783  =  339 bp
ORF:  112 amino acids  =  12.0 kDa
ORF:  4000 .. 4593  =  594 bp
ORF:  197 amino acids  =  20.5 kDa
ORF:  5362 .. 6129  =  768 bp
ORF:  255 amino acids  =  27.0 kDa
ORF:  6532 .. 6873  =  342 bp
ORF:  113 amino acids  =  12.6 kDa
ORF:  1371 .. 1688  =  318 bp
ORF:  105 amino acids  =  12.2 kDa
ORF:  2892 .. 3122  =  231 bp
ORF:  76 amino acids  =  8.4 kDa
ORF:  3789 .. 4142  =  354 bp
ORF:  117 amino acids  =  12.6 kDa
ORF:  5616 .. 5981  =  366 bp
ORF:  121 amino acids  =  13.3 kDa
ORF:  7299 .. 7562  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  8151 .. 8468  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  8720 .. 127  =  237 bp
ORF:  78 amino acids  =  8.5 kDa
ORF:  1403 .. 1681  =  279 bp
ORF:  92 amino acids  =  10.3 kDa
ORF:  6425 .. 6763  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
ORF:  7319 .. 8278  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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