pSMART HCAmp (linearized)

Linearized and dephosphorylated high-copy number vector with an ampicillin resistance marker for efficient blunt cloning of unstable sequences.

Sequence Author: Lucigen

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HomePlasmidsLucigen VectorspSMART HCAmp (linearized)
<HincII> (1833) EcoRI (1825) HindIII (1819) EcoRV (1815) SL1 (1775 .. 1795) AlwNI (1478) PspFI (1370) BseYI (1366) HaeII (1310) BciVI (1265) BspDI - ClaI (1162) ApaI - BanII (1068) EcoO109I - PspOMI (1064) BanI (1034) AhdI (987) BmrI (947) BsaI (921) <HincII> (0) EcoRI (4) XbaI (10) EcoRV (16) SwaI (37) SR2 (27 .. 52) BsaAI (106) Bpu10I (186) XmnI (387) BsaHI (447) TatI (504) ScaI (506) TsoI (589) EaeI (594) XcmI (604) PvuI (618) FspI (764) AseI (812) NmeAIII (840) BglI (869) BsrFI (902) pSMART® HCAmp 1833 bp
End  (1833)
0 sites
EcoRI  (1825)
2 sites		 G A A T T C C T T A A G
HindIII  (1819)
1 site		 A A G C T T T T C G A A
EcoRV  (1815)
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.
AlwNI  (1478)
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.
PspFI  (1370)
1 site		 C C C A G C G G G T C G
BseYI  (1366)
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  (1310)
1 site		 R G C G C Y Y C G C G R
BciVI  (1265)
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.
BspDI  (1162)
1 site		 A T C G A T T A G C T A
ClaI  (1162)
1 site		 A T C G A T T A G C T A
ApaI  (1068)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BanII  (1068)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
EcoO109I  (1064)
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  (1064)
1 site		 G G G C C C C C C G G G
BanI  (1034)
1 site		 G G Y R C C C C R Y G G
Sticky ends from different BanI sites may not be compatible.
AhdI  (987)
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.
BmrI  (947)
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.
BsaI  (921)
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.
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.
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
Bpu10I  (186)
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.
XmnI  (387)
1 site		 G A A N N N N T T C C T T N N N N A A G
BsaHI  (447)
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.
TatI  (504)
1 site		 W G T A C W W C A T G W
ScaI  (506)
1 site		 A G T A C T T C A T G A
TsoI  (589)
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).
EaeI  (594)
1 site		 Y G G C C R R C C G G Y
XcmI  (604)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
PvuI  (618)
1 site		 C G A T C G G C T A G C
FspI  (764)
1 site		 T G C G C A A C G C G T
AseI  (812)
1 site		 A T T A A T T A A T T A
NmeAIII  (840)
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).
BglI  (869)
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.
BsrFI  (902)
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.
SL1
21-mer  /  57% GC
1 binding site
1775 .. 1795  =  21 annealed bases
Tm  =  59°C
SR2
26-mer  /  38% GC
1 binding site
27 .. 52  =  26 annealed bases
Tm  =  56°C
AmpR
200 .. 1060  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   200 .. 268  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
200 .. 1060  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   269 .. 1060  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
200 .. 1060  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1123 .. 1710  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1123 .. 1710  =  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
1732 .. 1761  =  30 bp
phage T3 early transcription terminator
T3Te terminator
1732 .. 1761  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1084 .. 1111  =  28 bp
phage T7 early transcription terminator
T7Te terminator
1084 .. 1111  =  28 bp
phage T7 early transcription terminator
ORF:  200 .. 1060  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  664 .. 966  =  303 bp
ORF:  100 amino acids  =  10.5 kDa
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