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Plasmid Files

pSMART® HCKan

High-copy number vector with a kanamycin resistance marker for efficient blunt cloning of unstable sequences.

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pSMART HCKan Sequence and MappSMART HCKan.dna
Map and Sequence File   
Sequence Author:  Lucigen
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 EcoRI (1780) EcoRV (1770) BpmI (1764) SL1 (1730 .. 1750) AcuI (1565) AlwNI (1433) BsiHKAI (1335) ApaLI (1331) PspFI (1325) BseYI (1321) HaeII (1265) BciVI (1220) ApaI (1023) EcoO109I - PspOMI (1019) PflMI (894) HincII (0) EcoRI (4) XbaI (10) EcoRV (16) DraI - SwaI (37) SR2 (27 .. 52) BsaAI (106) MslI (129) AvaI - BsoBI - PaeR7I - PspXI - XhoI (229) BmeT110I (230) NruI (288) EarI (444) TsoI (472) PasI (503) EcoNI (543) SspI (556) BsrFI (585) AsiSI - PvuI (631) BsmBI (647) AseI (830) TaqII (885) pSMART® HCKan 1788 bp
EcoRI  (1780)
2 sites
G A A T T C C T T A A G
EcoRV  (1770)
2 sites
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.
BpmI  (1764)
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.
AcuI  (1565)
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).
AlwNI  (1433)
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.
BsiHKAI  (1335)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
ApaLI  (1331)
1 site
G T G C A C C A C G T G
PspFI  (1325)
1 site
C C C A G C G G G T C G
BseYI  (1321)
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.
HaeII  (1265)
1 site
R G C G C Y Y C G C G R
BciVI  (1220)
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.
ApaI  (1023)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoO109I  (1019)
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.
PspOMI  (1019)
1 site
G G G C C C C C C G G G
PflMI  (894)
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.
HincII  (0)
1 site
G T Y R A C C A R Y T G
EcoRI  (4)
2 sites
G A A T T C C T T A A G
XbaI  (10)
1 site
T C T A G A A G A T C T
EcoRV  (16)
2 sites
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.
DraI  (37)
1 site
T T T A A A A A A T T T
SwaI  (37)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
BsaAI  (106)
1 site
Y A C G T R R T G C A Y
MslI  (129)
1 site
C A Y N N N N R T G G T R N N N N Y A C
AvaI  (229)
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  (229)
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.
PaeR7I  (229)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (229)
1 site
V C T C G A G B B G A G C T C V
XhoI  (229)
1 site
C T C G A G G A G C T C
BmeT110I  (230)
1 site
C Y C G R G G R G C Y C
NruI  (288)
1 site
T C G C G A A G C G C T
EarI  (444)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
TsoI  (472)
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).
PasI  (503)
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.
EcoNI  (543)
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  (556)
1 site
A A T A T T T T A T A A
BsrFI  (585)
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  (631)
1 site
G C G A T C G C C G C T A G C G
PvuI  (631)
1 site
C G A T C G G C T A G C
BsmBI  (647)
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.
AseI  (830)
1 site
A T T A A T T A A T T A
TaqII  (885)
1 site
G A C C G A ( N ) 9 N N C T G G C T ( N ) 9

Sticky ends from different TaqII sites may not be compatible.
SL1
21-mer  /  57% GC
1 binding site
1730 .. 1750  =  21 annealed bases
Tm  =  59°C
SR2
26-mer  /  38% GC
1 binding site
27 .. 52  =  26 annealed bases
Tm  =  56°C
KanR
200 .. 1015  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
200 .. 1015  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
1078 .. 1665  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1078 .. 1665  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
cat promoter
97 .. 199  =  103 bp
promoter of the E. coli cat gene
cat promoter
97 .. 199  =  103 bp
promoter of the E. coli cat gene
tonB terminator
65 .. 96  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
tonB terminator
65 .. 96  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
T3Te terminator
1687 .. 1716  =  30 bp
phage T3 early transcription terminator
T3Te terminator
1687 .. 1716  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1039 .. 1066  =  28 bp
phage T7 early transcription terminator
T7Te terminator
1039 .. 1066  =  28 bp
phage T7 early transcription terminator
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