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Plasmid Files

pUC57-Kan

Cloning vector with a kanamycin resistance marker, suitable for generating ExoIII deletions.

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pUC57-Kan.dna
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Sequence Author:  GenScript
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BtgZI (2563) BsrDI (2544) EcoNI (2127) SspI (2115) BsrFI (2082) AsiSI (2042) Bpu10I (2020) PflMI (1780) AcuI - Eco57MI (1378) PfoI (46) TatI (167) NdeI (184) BstAPI (185) KasI (235) BsaHI - NarI (236) SfoI (237) PluTI (239) BglI (251) FspI (258) BmrI (359) EcoRI (396) Eco53kI (404) SacI (406) Acc65I (408) KpnI (412) 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) BssS α I (1003) BciVI (1033) BseYI (1134) PspFI (1138) AlwNI (1246) pUC57-Kan 2579 bp
BtgZI  (2563)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsrDI  (2544)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
EcoNI  (2127)
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.
SspI  (2115)
1 site
A A T A T T T T A T A A
BsrFI  (2082)
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.
AsiSI  (2042)
1 site
G C G A T C G C C G C T A G C G
Bpu10I  (2020)
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.
PflMI  (1780)
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.
AcuI  (1378)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
Eco57MI  (1378)
1 site
C T G R A G ( N ) 14 N N G A C Y T C ( N ) 14

Sticky ends from different Eco57MI sites may not be compatible.
After cleavage, Eco57MI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
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.
TatI  (167)
1 site
W G T A C W W C A T G W
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
BsaHI  (236)
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.
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.
BglI  (251)
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.
FspI  (258)
1 site
T G C G C A A C G C G T
BmrI  (359)
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.
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
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.
BssSαI  (1003)
1 site
C A C G A G G T G C T C
BciVI  (1033)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
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.
KanR
1657 .. 2466  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
1657 .. 2466  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
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
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
ORF:  186 .. 566  =  381 bp
ORF:  126 amino acids  =  14.5 kDa
ORF:  2514 .. 174  =  240 bp
ORF:  79 amino acids  =  9.0 kDa
ORF:  146 .. 493  =  348 bp
ORF:  115 amino acids  =  13.1 kDa
ORF:  1657 .. 2466  =  810 bp
ORF:  269 amino acids  =  30.8 kDa
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