pSMART LCKan (linearized)

Linearized and dephosphorylated low-copy number vector with a kanamycin resistance marker for efficient blunt cloning of unstable sequences.

Sequence Author: Lucigen

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HomePlasmidsLucigen VectorspSMART LCKan (linearized)
<HincII> (1968) EcoRI (1960) EcoRV (1950) SL1 (1910 .. 1930) AcuI (1745) AlwNI (1613) BsiHKAI (1515) ApaLI (1511) PspFI (1505) BseYI (1501) HaeII (1445) BciVI (1400) ApaI (1203) EcoO109I - PspOMI (1199) PvuII (1155) XmnI (1122) <HincII> (0) EcoRI (4) XbaI (10) EcoRV (16) DraI - SwaI (37) SR2 (27 .. 52) BsaAI (106) AvaI - BmeT110I - BsoBI - PaeR7I - PspXI - XhoI (217) NruI (276) EarI (432) TsoI (460) PasI (491) EcoNI (531) SspI (544) BsrFI (573) AsiSI - PvuI (619) BsmBI (635) Bpu10I (636) AseI (818) TaqII (873) PflMI (882) pSMART® LCKan 1968 bp
End  (1968)
0 sites
EcoRI  (1960)
2 sites		 G A A T T C C T T A A G
EcoRV  (1950)
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.
AcuI  (1745)
1 site		 C T G A A G ( N ) 14 N N G A C T T C ( N ) 14
Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
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  (1613)
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  (1515)
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  (1511)
1 site		 G T G C A C C A C G T G
PspFI  (1505)
1 site		 C C C A G C G G G T C G
BseYI  (1501)
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  (1445)
1 site		 R G C G C Y Y C G C G R
BciVI  (1400)
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  (1203)
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  (1199)
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  (1199)
1 site		 G G G C C C C C C G G G
PvuII  (1155)
1 site		 C A G C T G G T C G A C
XmnI  (1122)
1 site		 G A A N N N N T T C C T T N N N N A A G
Start  (0)
0 sites
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
AvaI  (217)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BmeT110I  (217)
1 site		 C Y C G R G G R G C Y C
BsoBI  (217)
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  (217)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (217)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (217)
1 site		 C T C G A G G A G C T C
NruI  (276)
1 site		 T C G C G A A G C G C T
EarI  (432)
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.
TsoI  (460)
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  (491)
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  (531)
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  (544)
1 site		 A A T A T T T T A T A A
BsrFI  (573)
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.
AsiSI  (619)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (619)
1 site		 C G A T C G G C T A G C
BsmBI  (635)
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.
Bpu10I  (636)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
AseI  (818)
1 site		 A T T A A T T A A T T A
TaqII  (873)
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.
PflMI  (882)
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.
SL1
21-mer  /  57% GC
1 binding site
1910 .. 1930  =  21 annealed bases
Tm  =  59°C
SR2
26-mer  /  38% GC
1 binding site
27 .. 52  =  26 annealed bases
Tm  =  56°C
KanR
188 .. 1003  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
188 .. 1003  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
1258 .. 1845  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1258 .. 1845  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
1004 .. 1195  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
1004 .. 1195  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
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
1867 .. 1896  =  30 bp
phage T3 early transcription terminator
T3Te terminator
1867 .. 1896  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1219 .. 1246  =  28 bp
phage T7 early transcription terminator
T7Te terminator
1219 .. 1246  =  28 bp
phage T7 early transcription terminator
ORF:  188 .. 1003  =  816 bp
ORF:  271 amino acids  =  30.9 kDa
ORF:  1098 .. 1583  =  486 bp
ORF:  161 amino acids  =  18.0 kDa
ORF:  1061 .. 1288  =  228 bp
ORF:  75 amino acids  =  8.6 kDa
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Individual Sequences & Maps

pAviTag C-His Kan
pEZ BAC
pRANGER-BTB-2
pSMART GC HK (linearized)
pAviTag N-His Kan
pEZ BAC (BamHI linearized)
pRANGER-BTB-3
pSMART GC LK (linearized)
pAviTag-C N-His SUMO Kan
pEZ BAC (HpaI linearized)
pRANGER-BTB-5
pSMART HCAmp
pAviTag-N N-His SUMO Kan
pGC Blue (linearized)
pRham C-His Kan
pSMART HCAmp (linearized)
pETite C-His Kan
pInvRecA
pRham N-His Kan
pSMART HCKan
pETite N-His Kan
pJAZZ-OC
pRham N-His SUMO Kan
pSMART HCKan (linearized)
pETite N-His SUMO Kan
pJAZZ-OK
pSMART BAC v2.0
pSMART LCAmp
pEZSeq-Amp
pJW168
pSMART BAC v2.0 (BamHI linearized)
pSMART LCAmp (linearized)
pEZSeq-Amp (linearized)
pME-HA (linearized)
pSMART BAC v2.0 (EcoRI linearized)
pSMART LCKan
pEZSeq-Kan
pNGS FOS (linearized)
pSMART BAC v2.0 (HindIII linearized)
pSMART LCKan (linearized)
pEZSeq-Kan (linearized)
pRANGER-BTB-1
pSMART FOS v2.0 (linearized)

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