pLUG-Multi (linearized)

Linearized TA cloning vector with 3'-T overhangs and a symmetric multiple cloning site for cloning of PCR products.

Sequence Author: iNtRON Biotechnology

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HomePlasmidsTA and GC Cloning VectorspLUG-Multi (linearized)
BspQI - SapI (2463) AflIII - PciI (2346) PspFI (2046) BseYI (2042) AlwNI (1937) AhdI (1458) BsaI (1392) BpmI (1389) BsrFI (1373) NmeAIII (1311) TsoI (1060) EcoRI (2702) SacI (2712) KpnI (2718) SmaI (2720) BamHI (2723) XbaI (2729) SalI (2735) PstI (2745) SphI (2751) HindIII (2753) End (2766) Start (1) BmgBI (4) HindIII (10) SphI (20) PstI (26) SalI (28) XbaI (34) BamHI (40) SmaI (47) KpnI (53) SacI (59) EcoRI (61) KasI (231) NarI (232) SfoI (233) BbeI (235) NdeI (284) BstAPI (288) PfoI (419) EcoO109I (478) ZraI (537) AatII (539) SspI (653) XmnI (858) ScaI (977) pLUG®-Multi 2765 bp
BspQI  (2463)
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  (2463)
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  (2346)
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  (2346)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PspFI  (2046)
1 site		 C C C A G C G G G T C G
BseYI  (2042)
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.
AlwNI  (1937)
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.
AhdI  (1458)
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.
BsaI  (1392)
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.
BpmI  (1389)
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.
BsrFI  (1373)
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.
NmeAIII  (1311)
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).
TsoI  (1060)
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).
EcoRI  (2702)
2 sites		 G A A T T C C T T A A G
SacI  (2712)
2 sites		 G A G C T C C T C G A G
KpnI  (2718)
2 sites		 G G T A C C C C A T G G
SmaI  (2720)
2 sites		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BamHI  (2723)
2 sites		 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.
XbaI  (2729)
2 sites		 T C T A G A A G A T C T
SalI  (2735)
2 sites		 G T C G A C C A G C T G
PstI  (2745)
2 sites		 C T G C A G G A C G T C
SphI  (2751)
2 sites		 G C A T G C C G T A C G
HindIII  (2753)
2 sites		 A A G C T T T T C G A A
End  (2766)
0 sites
Start  (1)
0 sites
BmgBI  (4)
1 site		 C A C G T C G T G C A G
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
HindIII  (10)
2 sites		 A A G C T T T T C G A A
SphI  (20)
2 sites		 G C A T G C C G T A C G
PstI  (26)
2 sites		 C T G C A G G A C G T C
SalI  (28)
2 sites		 G T C G A C C A G C T G
XbaI  (34)
2 sites		 T C T A G A A G A T C T
BamHI  (40)
2 sites		 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.
SmaI  (47)
2 sites		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
KpnI  (53)
2 sites		 G G T A C C C C A T G G
SacI  (59)
2 sites		 G A G C T C C T C G A G
EcoRI  (61)
2 sites		 G A A T T C C T T A A G
KasI  (231)
1 site		 G G C G C C C C G C G G
NarI  (232)
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  (233)
1 site		 G G C G C C C C G C G G
BbeI  (235)
1 site		 G G C G C C C C G C G G
Cleavage may be enhanced when more than one copy of the BbeI recognition sequence is present.
NdeI  (284)
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  (288)
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.
PfoI  (419)
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.
EcoO109I  (478)
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.
ZraI  (537)
1 site		 G A C G T C C T G C A G
AatII  (539)
1 site		 G A C G T C C T G C A G
SspI  (653)
1 site		 A A T A T T T T A T A A
XmnI  (858)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (977)
1 site		 A G T A C T T C A T G A
AmpR
671 .. 1531  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   671 .. 739  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
671 .. 1531  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   740 .. 1531  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
671 .. 1531  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1702 .. 2290  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1702 .. 2290  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
lacZα
2 .. 325  =  324 bp
107 amino acids  =  12.1 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2 .. 325  =  324 bp
107 amino acids  =  12.1 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
566 .. 670  =  105 bp
AmpR promoter
566 .. 670  =  105 bp
lacZα
2688 .. 2765  =  78 bp
26 amino acids  =  2.8 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2688 .. 2765  =  78 bp
26 amino acids  =  2.8 kDa
Product: LacZα fragment of β-galactosidase
lac promoter
2614 .. 2644  =  31 bp
3 segments
   Segment 1:  -35  
   2614 .. 2619  =  6 bp
promoter for the E. coli lac operon
lac promoter
2614 .. 2644  =  31 bp
3 segments
   Segment 2:  
   2620 .. 2637  =  18 bp
promoter for the E. coli lac operon
lac promoter
2614 .. 2644  =  31 bp
3 segments
   Segment 3:  -10  
   2638 .. 2644  =  7 bp
promoter for the E. coli lac operon
lac promoter
2614 .. 2644  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
2652 .. 2668  =  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
2652 .. 2668  =  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
10 .. 66  =  57 bp
pUC18/19 multiple cloning site
MCS
10 .. 66  =  57 bp
pUC18/19 multiple cloning site
MCS
2702 .. 2758  =  57 bp
pUC18/19 multiple cloning site
MCS
2702 .. 2758  =  57 bp
pUC18/19 multiple cloning site
M13 fwd
76 .. 92  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
76 .. 92  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
2676 .. 2692  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
2676 .. 2692  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  2 .. 325  =  324 bp
ORF:  107 amino acids  =  12.1 kDa  (no start codon)
ORF:  671 .. 1531  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1 .. 285  =  285 bp
ORF:  95 amino acids  =  10.9 kDa
ORF:  1135 .. 1401  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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