pUCmu

Minimal 1669-bp cloning vector derived from pUC18, with an ampicillin resistance marker. See also pICOz.

Sequence Author: BCCM/GeneCorner Plasmid Collection

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NdeI (16) NruI (8) MluI (1) BsrBI (1659) BspHI (1653) SspI (1622) EarI (1613) XmnI (1417) BsaHI (1355) ScaI (1298) TatI (1296) TsoI (1217) PvuI (1188) FspI (1040) AseI (990) NmeAIII (966) BglI (938) BsrFI (898) BpmI (888) BsaI (879) BmrI (858) HpaI (24) BtgI - NcoI - StyI (28) MscI (33) HindIII (36) BfuAI - BspMI (41) SphI (46) PstI - SbfI (52) SalI (54) AccI (55) XbaI (60) BamHI (66) TspMI - XmaI (70) SmaI (72) Acc65I (74) KpnI (78) Eco53kI (82) SacI (84) ApoI - EcoRI (86) EcoRV (95) PaeR7I - XhoI (99) SpeI (105) PspOMI (111) ApaI (115) PmeI (120) PciI (125) DrdI (181) HaeII (321) BseYI (377) PspFI (381) AlwNI (489) AhdI (818) pUCmu 1669 bp
NdeI  (16)
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.
NruI  (8)
1 site
T C G C G A A G C G C T
MluI  (1)
1 site
A C G C G T T G C G C A
BsrBI  (1659)
1 site
C C G C T C G G C G A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures up to 50°C.
BspHI  (1653)
1 site
T C A T G A A G T A C T
SspI  (1622)
1 site
A A T A T T T T A T A A
EarI  (1613)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
XmnI  (1417)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (1355)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
ScaI  (1298)
1 site
A G T A C T T C A T G A
TatI  (1296)
1 site
W G T A C W W C A T G W
TsoI  (1217)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PvuI  (1188)
1 site
C G A T C G G C T A G C
FspI  (1040)
1 site
T G C G C A A C G C G T
AseI  (990)
1 site
A T T A A T T A A T T A
NmeAIII  (966)
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).
BglI  (938)
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.
BsrFI  (898)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
BpmI  (888)
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.
BsaI  (879)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BmrI  (858)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
HpaI  (24)
1 site
G T T A A C C A A T T G
BtgI  (28)
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  (28)
1 site
C C A T G G G G T A C C
StyI  (28)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MscI  (33)
1 site
T G G C C A A C C G G T
HindIII  (36)
1 site
A A G C T T T T C G A A
BfuAI  (41)
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  (41)
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.
SphI  (46)
1 site
G C A T G C C G T A C G
PstI  (52)
1 site
C T G C A G G A C G T C
SbfI  (52)
1 site
C C T G C A G G G G A C G T C C
SalI  (54)
1 site
G T C G A C C A G C T G
AccI  (55)
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.
XbaI  (60)
1 site
T C T A G A A G A T C T
BamHI  (66)
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  (70)
1 site
C C C G G G G G G C C C
XmaI  (70)
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.
SmaI  (72)
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  (74)
1 site
G G T A C C C C A T G G
KpnI  (78)
1 site
G G T A C C C C A T G G
Eco53kI  (82)
1 site
G A G C T C C T C G A G
SacI  (84)
1 site
G A G C T C C T C G A G
ApoI  (86)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (86)
1 site
G A A T T C C T T A A G
EcoRV  (95)
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.
PaeR7I  (99)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (99)
1 site
C T C G A G G A G C T C
SpeI  (105)
1 site
A C T A G T T G A T C A
PspOMI  (111)
1 site
G G G C C C C C C G G G
ApaI  (115)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PmeI  (120)
1 site
G T T T A A A C C A A A T T T G
PciI  (125)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (181)
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.
HaeII  (321)
1 site
R G C G C Y Y C G C G R
BseYI  (377)
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.
PspFI  (381)
1 site
C C C A G C G G G T C G
AlwNI  (489)
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.
AhdI  (818)
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.
AmpR
745 .. 1605  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   745 .. 1536  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
745 .. 1605  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1537 .. 1605  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
745 .. 1605  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
134 .. 722  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
134 .. 722  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
MCS
1 .. 130  =  130 bp
multiple cloning site
MCS
1 .. 130  =  130 bp
multiple cloning site
AmpR promoter
1606 .. 1669  =  64 bp
AmpR promoter
1606 .. 1669  =  64 bp
ORF:  875 .. 1141  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1655 .. 459  =  474 bp
ORF:  157 amino acids  =  17.6 kDa
ORF:  745 .. 1605  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

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