pSMART GC LK (linearized)

Linearized and dephosphorylated low copy-number vector with 3'-C tails for transcription-free GC cloning of PCR products.

Sequence Author: Lucigen

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HomePlasmidsTA and GC Cloning VectorspSMART GC LK (linearized)
End (1993) EcoRI (1979) EcoRV (1969) SL1 Primer (1929 .. 1949) AcuI (1764) AlwNI (1632) BsiHKAI (1534) ApaLI (1530) PspFI (1524) BseYI (1520) HaeII (1464) BciVI (1419) ApaI (1222) EcoO109I - PspOMI (1218) PvuII (1174) XmnI (1141) Start (1) EcoRI (10) XbaI (16) EcoRV (22) DraI - SwaI (43) SR2 Primer (33 .. 58) BsaAI (112) AvaI - BmeT110I - BsoBI - PaeR7I - PspXI - XhoI (235) NruI (294) EarI (450) TsoI (478) PasI (509) EcoNI (549) SspI (562) BsrFI (591) AsiSI - PvuI (637) BsmBI (653) AseI (836) TaqII (891) PflMI (900) pSMART® GC LK 1992 bp
End  (1993)
0 sites
EcoRI  (1979)
2 sites		 G A A T T C C T T A A G
EcoRV  (1969)
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  (1764)
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  (1632)
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  (1534)
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  (1530)
1 site		 G T G C A C C A C G T G
PspFI  (1524)
1 site		 C C C A G C G G G T C G
BseYI  (1520)
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  (1464)
1 site		 R G C G C Y Y C G C G R
BciVI  (1419)
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  (1222)
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  (1218)
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  (1218)
1 site		 G G G C C C C C C G G G
PvuII  (1174)
1 site		 C A G C T G G T C G A C
XmnI  (1141)
1 site		 G A A N N N N T T C C T T N N N N A A G
Start  (1)
0 sites
EcoRI  (10)
2 sites		 G A A T T C C T T A A G
XbaI  (16)
1 site		 T C T A G A A G A T C T
EcoRV  (22)
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  (43)
1 site		 T T T A A A A A A T T T
SwaI  (43)
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  (112)
1 site		 Y A C G T R R T G C A Y
AvaI  (235)
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  (235)
1 site		 C Y C G R G G R G C Y C
BsoBI  (235)
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  (235)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (235)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (235)
1 site		 C T C G A G G A G C T C
NruI  (294)
1 site		 T C G C G A A G C G C T
EarI  (450)
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  (478)
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  (509)
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  (549)
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  (562)
1 site		 A A T A T T T T A T A A
BsrFI  (591)
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  (637)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (637)
1 site		 C G A T C G G C T A G C
BsmBI  (653)
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.
AseI  (836)
1 site		 A T T A A T T A A T T A
TaqII  (891)
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  (900)
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 Primer
21-mer  /  57% GC
1 binding site
1929 .. 1949  =  21 annealed bases
Tm  =  59°C
SR2 Primer
26-mer  /  38% GC
1 binding site
33 .. 58  =  26 annealed bases
Tm  =  56°C
KanR
206 .. 1021  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
206 .. 1021  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
1277 .. 1864  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1277 .. 1864  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
1026 .. 1214  =  189 bp
62 amino acids  =  7.1 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
1026 .. 1214  =  189 bp
62 amino acids  =  7.1 kDa
Product: Rop protein, which maintains plasmids at low copy number
cat promoter
103 .. 205  =  103 bp
promoter of the E. coli cat gene
cat promoter
103 .. 205  =  103 bp
promoter of the E. coli cat gene
tonB terminator
71 .. 102  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
tonB terminator
71 .. 102  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
T3Te terminator
1886 .. 1915  =  30 bp
phage T3 early transcription terminator
T3Te terminator
1886 .. 1915  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1238 .. 1265  =  28 bp
phage T7 early transcription terminator
T7Te terminator
1238 .. 1265  =  28 bp
phage T7 early transcription terminator
ORF:  1117 .. 1602  =  486 bp
ORF:  161 amino acids  =  18.0 kDa
ORF:  206 .. 1021  =  816 bp
ORF:  271 amino acids  =  30.9 kDa
ORF:  1080 .. 1307  =  228 bp
ORF:  75 amino acids  =  8.6 kDa
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Individual Sequences & Maps

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pLUG (linearized)
pTargeT (linearized)
pcDNA3.1 V5-His-TOPO (linearized)
pENTR5'-TOPO (linearized)
pLUG-Multi (linearized)
pTOP TA V2 (linearized)
pcDNA3.3-TOPO (linearized)
pET SUMO (linearized)
pMT V5-His-TOPO (linearized)
pTrcHis-TOPO (linearized)
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pETBlue-1 AccepTor
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pEXP5-CT TOPO (linearized)
pPrime (linearized)
pTZ57R_T
pcDNA5 FRT TO-TOPO (linearized)
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pcDNA5 FRT V5-His-TOPO (linearized)
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pSC-A-amp kan (linearized)
pYES2.1 V5-His-TOPO (linearized)
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T-Vector pMD19 (Simple)
pcDNA6.2 N-EmGFP-GW TOPO (linearized)
pGlow-TOPO (linearized)
pSMART GC LK (linearized)
T-Vector pMD20
pCR-XL-TOPO (linearized)
pIB V5-His-TOPO (linearized)

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