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Plasmid Files

pUC57

Cloning vector with an ampicillin resistance marker, suitable for generating ExoIII deletions.

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pUC57 Sequence and MappUC57.dna
Map and Sequence File   
Sequence Author:  GenScript
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 EcoO109I (2699) AatII (2645) ZraI (2643) SspI (2527) XmnI (2322) ScaI (2203) TsoI (2122) NmeAIII (1871) BsrFI (1803) BpmI (1793) BsaI (1784) AhdI (1723) PfoI (46) NdeI (184) BstAPI (185) KasI (235) NarI (236) SfoI (237) PluTI (239) ApoI - EcoRI (396) Eco53kI (404) SacI (406) Acc65I (408) KpnI (412) NruI (416) BsmI - NsiI (424) XbaI (425) EcoRV (431) BamHI (435) AvaI - BsoBI - TspMI - XmaI (439) BmeT110I (440) SmaI (441) PspOMI (442) ApaI (446) SalI (448) AccI (449) HincII (450) PstI (457) StuI (461) SphI (469) HindIII (471) BspQI - SapI (714) AflIII - PciI (830) BseYI (1134) PspFI (1138) AlwNI (1246) pUC57 2710 bp
EcoO109I  (2699)
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.
AatII  (2645)
1 site
G A C G T C C T G C A G
ZraI  (2643)
1 site
G A C G T C C T G C A G
SspI  (2527)
1 site
A A T A T T T T A T A A
XmnI  (2322)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2203)
1 site
A G T A C T T C A T G A
TsoI  (2122)
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).
NmeAIII  (1871)
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).
BsrFI  (1803)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI  (1793)
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  (1784)
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.
AhdI  (1723)
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.
PfoI  (46)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
NdeI  (184)
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.
BstAPI  (185)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
KasI  (235)
1 site
G G C G C C C C G C G G
NarI  (236)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (237)
1 site
G G C G C C C C G C G G
PluTI  (239)
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.
ApoI  (396)
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  (396)
1 site
G A A T T C C T T A A G
Eco53kI  (404)
1 site
G A G C T C C T C G A G
SacI  (406)
1 site
G A G C T C C T C G A G
Acc65I  (408)
1 site
G G T A C C C C A T G G
KpnI  (412)
1 site
G G T A C C C C A T G G
NruI  (416)
1 site
T C G C G A A G C G C T
BsmI  (424)
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.
NsiI  (424)
1 site
A T G C A T T A C G T A
XbaI  (425)
1 site
T C T A G A A G A T C T
EcoRV  (431)
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  (435)
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.
AvaI  (439)
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  (439)
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.
TspMI  (439)
1 site
C C C G G G G G G C C C
XmaI  (439)
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.
BmeT110I  (440)
1 site
C Y C G R G G R G C Y C
SmaI  (441)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PspOMI  (442)
1 site
G G G C C C C C C G G G
ApaI  (446)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SalI  (448)
1 site
G T C G A C C A G C T G
AccI  (449)
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.
HincII  (450)
1 site
G T Y R A C C A R Y T G
PstI  (457)
1 site
C T G C A G G A C G T C
StuI  (461)
1 site
A G G C C T T C C G G A
SphI  (469)
1 site
G C A T G C C G T A C G
HindIII  (471)
1 site
A A G C T T T T C G A A
BspQI  (714)
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  (714)
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.
AflIII  (830)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (830)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (1134)
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  (1138)
1 site
C C C A G C G G G T C G
AlwNI  (1246)
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.
AmpR
1650 .. 2510  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1650 .. 2441  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1650 .. 2510  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2442 .. 2510  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1650 .. 2510  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
891 .. 1479  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
891 .. 1479  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lacZα
146 .. 493  =  348 bp
115 amino acids  =  13.1 kDa
Product: LacZα fragment of β-galactosidase
lacZα
146 .. 493  =  348 bp
115 amino acids  =  13.1 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
2511 .. 2615  =  105 bp
AmpR promoter
2511 .. 2615  =  105 bp
lac promoter
537 .. 567  =  31 bp
   Segment 3:  -10  
   537 .. 543  =  7 bp
promoter for the E. coli lac operon
lac promoter
537 .. 567  =  31 bp
   Segment 2:  
   544 .. 561  =  18 bp
promoter for the E. coli lac operon
lac promoter
537 .. 567  =  31 bp
   Segment 1:  -35  
   562 .. 567  =  6 bp
promoter for the E. coli lac operon
lac promoter
537 .. 567  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
513 .. 529  =  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
513 .. 529  =  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).
MCS
396 .. 476  =  81 bp
multiple cloning site
MCS
396 .. 476  =  81 bp
multiple cloning site
M13 fwd
379 .. 395  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
379 .. 395  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
489 .. 505  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
489 .. 505  =  17 bp
common sequencing primer, one of multiple similar
variants
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