Resources
Plasmid Files

pMiniT

Compact bacterial vector that employs a toxic minigene for high-efficiency cloning of PCR products.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pMiniT Sequence and MappMiniT.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
Download Free Trial Get SnapGene Viewer

 BspQI - SapI (2462) NspI (2349) AflIII - PciI (2345) DrdI (2243) PspFI (2045) BseYI (2041) AlwNI (1936) BanI (1504) AhdI (1457) BmrI (1417) BsaI (1391) BpmI (1388) BglI (1339) NmeAIII (1310) BtgZI (99) AfeI (143) NdeI (207) NruI (225) tnpA transposase Cloning Analysis Forward Primer (391 .. 412) BfuAI - BspMI (400) PspXI - XhoI (486) BamHI (492) EcoRI (498) Shine-Dalgarno sequence toxic minigene ZraI (536) AatII (538) EcoRI (541) XhoI (546) Cloning Analysis Reverse Primer (614 .. 636) SspI (652) XmnI (857) TatI (974) ScaI (976) TsoI (1059) PvuI (1088) FspI (1234) pMiniT 2525 bp
BspQI  (2462)
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  (2462)
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.
NspI  (2349)
1 site
R C A T G Y Y G T A C R
AflIII  (2345)
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  (2345)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (2243)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PspFI  (2045)
1 site
C C C A G C G G G T C G
BseYI  (2041)
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  (1936)
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.
BanI  (1504)
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  (1457)
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  (1417)
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  (1391)
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  (1388)
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.
BglI  (1339)
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.
NmeAIII  (1310)
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).
BtgZI  (99)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
AfeI  (143)
1 site
A G C G C T T C G C G A
NdeI  (207)
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.
NruI  (225)
1 site
T C G C G A A G C G C T
BfuAI  (400)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (400)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PspXI  (486)
1 site
V C T C G A G B B G A G C T C V
XhoI  (486)
2 sites
C T C G A G G A G C T C
BamHI  (492)
1 site
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.
EcoRI  (498)
2 sites
G A A T T C C T T A A G
ZraI  (536)
1 site
G A C G T C C T G C A G
AatII  (538)
1 site
G A C G T C C T G C A G
EcoRI  (541)
2 sites
G A A T T C C T T A A G
XhoI  (546)
2 sites
C T C G A G G A G C T C
SspI  (652)
1 site
A A T A T T T T A T A A
XmnI  (857)
1 site
G A A N N N N T T C C T T N N N N A A G
TatI  (974)
1 site
W G T A C W W C A T G W
ScaI  (976)
1 site
A G T A C T T C A T G A
TsoI  (1059)
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).
PvuI  (1088)
1 site
C G A T C G G C T A G C
FspI  (1234)
1 site
T G C G C A A C G C G T
Cloning Analysis Forward Primer
22-mer  /  55% GC
1 binding site
391 .. 412  =  22 annealed bases
Tm  =  62°C
Cloning Analysis Reverse Primer
23-mer  /  48% GC
1 binding site
614 .. 636  =  23 annealed bases
Tm  =  57°C
AmpR
670 .. 1530  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   670 .. 738  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
670 .. 1530  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   739 .. 1530  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
670 .. 1530  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1701 .. 2289  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1701 .. 2289  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
tnpA transposase
252 .. 482  =  231 bp
77 amino acids  =  9.0 kDa
Product: transposase encoded by the IS10 tnpA
gene
tnpA transposase
252 .. 482  =  231 bp
77 amino acids  =  9.0 kDa
Product: transposase encoded by the IS10 tnpA
gene
tnpA promoter
145 .. 251  =  107 bp
promoter of the IS10 transposase
tnpA promoter
145 .. 251  =  107 bp
promoter of the IS10 transposase
AmpR promoter
565 .. 669  =  105 bp
AmpR promoter
565 .. 669  =  105 bp
stop codons
524 .. 535  =  12 bp
stop codons
524 .. 535  =  12 bp
Shine-Dalgarno sequence
504 .. 510  =  7 bp
ribosome binding site
Shine-Dalgarno sequence
504 .. 510  =  7 bp
ribosome binding site
toxic minigene
518 .. 523  =  6 bp
2 amino acids  =  262.4 Da
Product: two-residue polypeptide that poisons the E.
coli
translation machinery
toxic minigene
518 .. 523  =  6 bp
2 amino acids  =  262.4 Da
Product: two-residue polypeptide that poisons the E.
coli
translation machinery
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter