pMiniT 2.0 (linearized)

Linearized compact bacterial vector that employs a toxic minigene for high-efficiency cloning and subsequent in vitro transcription of PCR products.

Sequence Author: New England Biolabs

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HomePlasmidsBasic Cloning VectorspMiniT 2.0 (linearized)
BspQI - SapI (1980) AflIII - PciI (1863) DrdI (1761) PspFI (1563) BseYI (1559) AlwNI (1454) BanI (1022) AhdI (975) BmrI (935) BpmI (906) BglI (857) NmeAIII (828) FspI (752) BtgZI (2142) AfeI (2186) NdeI (2250) NruI (2268) Cloning Analysis Forward Primer (2434 .. 2455) PstI - SbfI (2506) AarI (2509) PmeI (2515) PspXI - XhoI (2552) BamHI (2558) EcoRI (2564) Shine-Dalgarno sequence toxic minigene End (2588) Start (0) toxic minigene PacI (15) ZraI (21) AatII (23) EcoRI (26) XhoI (31) EagI - NotI (38) BsmBI (56) Cloning Analysis Reverse Primer (132 .. 154) SspI (170) XmnI (375) TatI (492) ScaI (494) TsoI (577) PvuI (606) pMiniT 2.0 2588 bp
BspQI  (1980)
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  (1980)
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  (1863)
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  (1863)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DrdI  (1761)
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  (1563)
1 site		 C C C A G C G G G T C G
BseYI  (1559)
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  (1454)
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  (1022)
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  (975)
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  (935)
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.
BpmI  (906)
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  (857)
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  (828)
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).
FspI  (752)
1 site		 T G C G C A A C G C G T
BtgZI  (2142)
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  (2186)
1 site		 A G C G C T T C G C G A
NdeI  (2250)
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  (2268)
1 site		 T C G C G A A G C G C T
PstI  (2506)
1 site		 C T G C A G G A C G T C
SbfI  (2506)
1 site		 C C T G C A G G G G A C G T C C
AarI  (2509)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
PmeI  (2515)
1 site		 G T T T A A A C C A A A T T T G
PspXI  (2552)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2552)
2 sites		 C T C G A G G A G C T C
BamHI  (2558)
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  (2564)
2 sites		 G A A T T C C T T A A G
End  (2588)
0 sites
Start  (0)
0 sites
PacI  (15)
1 site		 T T A A T T A A A A T T A A T T
ZraI  (21)
1 site		 G A C G T C C T G C A G
AatII  (23)
1 site		 G A C G T C C T G C A G
EcoRI  (26)
2 sites		 G A A T T C C T T A A G
XhoI  (31)
2 sites		 C T C G A G G A G C T C
EagI  (38)
1 site		 C G G C C G G C C G G C
NotI  (38)
1 site		 G C G G C C G C C G C C G G C G
BsmBI  (56)
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.
SspI  (170)
1 site		 A A T A T T T T A T A A
XmnI  (375)
1 site		 G A A N N N N T T C C T T N N N N A A G
TatI  (492)
1 site		 W G T A C W W C A T G W
ScaI  (494)
1 site		 A G T A C T T C A T G A
TsoI  (577)
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  (606)
1 site		 C G A T C G G C T A G C
Cloning Analysis Forward Primer
22-mer  /  55% GC
1 binding site
2434 .. 2455  =  22 annealed bases
Tm  =  62°C
Cloning Analysis Reverse Primer
23-mer  /  48% GC
1 binding site
132 .. 154  =  23 annealed bases
Tm  =  59°C
AmpR
188 .. 1048  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   188 .. 256  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
188 .. 1048  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   257 .. 1048  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
188 .. 1048  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1219 .. 1807  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1219 .. 1807  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
tnpA promoter
2188 .. 2294  =  107 bp
promoter of the IS10 transposase
tnpA promoter
2188 .. 2294  =  107 bp
promoter of the IS10 transposase
AmpR promoter
84 .. 187  =  104 bp
AmpR promoter
84 .. 187  =  104 bp
T7 promoter
56 .. 74  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
56 .. 74  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
2522 .. 2540  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
2522 .. 2540  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
stop codons
2 .. 10  =  9 bp
stop codons
2 .. 10  =  9 bp
Shine-Dalgarno sequence
2570 .. 2576  =  7 bp
ribosome binding site
Shine-Dalgarno sequence
2570 .. 2576  =  7 bp
ribosome binding site
toxic minigene
2584 .. 2588  =  5 bp
1 amino acid  =  149.2 Da
Product: two-residue polypeptide that poisons the E. coli translation machinery
toxic minigene
2584 .. 2588  =  5 bp
1 amino acid  =  149.2 Da
Product: two-residue polypeptide that poisons the E. coli translation machinery
toxic minigene
1 .. 1  =  1 bp
0 codons
Product: two-residue polypeptide that poisons the E. coli translation machinery
toxic minigene
1 .. 1  =  1 bp
0 codons
Product: two-residue polypeptide that poisons the E. coli translation machinery
ORF:  188 .. 1048  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2295 .. 2531  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  652 .. 918  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pMiniT 2.0 (linearized).dna file

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Individual Sequences & Maps

BlueScribe
pEASY-T1 (linearized)
pJET1.2
pSMART LCAmp
CAG promoter
pEASY-T1 Simple (linearized)
pJET1.2 blunt
pSMART LCAmp (linearized)
chicken beta-actin promoter
pEASY-T3 (linearized)
pJW168
pSMART LCKan
CLIP-tag
pEASY-T5 Zero (linearized)
pKF 18k-2
pSMART LCKan (linearized)
CMV promoter
pEX-1
pKF 19k-2
pSP64 Poly(A)
GAL1 promoter
pEX-A
pKOV
pSP72
GST
pEX-C-GST
pLATE11 (linearized)
pSP73
HaloTag
pEX-C-His
pLATE31 (linearized)
pSpark Done (linearized)
HSV TK promoter
pEX-K
pLATE51 (linearized)
pSpark I (linearized)
lac operator
pEX-N-GST
pLATE52 (linearized)
pSpark II (linearized)
lac promoter
pEX-N-His
pLEXX-AK
pSpark III (linearized)
lac UV5 promoter
pEX-N-His-GST
pLPS-B (linearized)
pSpark IV (linearized)
lacZ
pEX1
pMAL-c5E
pSpark V (linearized)
lambda
pEX2
pMAL-c5G
pSpot1
LITMUS28
pEX3
pMAL-c5X
pSpot2
LITMUS28i
pEZSeq-Amp
pMAL-c5X-His
pSpot3
LITMUS29
pEZSeq-Amp (linearized)
pMAL-c6T
pSpot4
LITMUS38
pEZSeq-Kan
pMAL-p5E
pSTBlue-1
LITMUS38i
pEZSeq-Kan (linearized)
pMAL-p5G
pSTV28
LITMUS39
pEZ BAC
pMAL-p5X
pSTV29
M13mp18
pEZ BAC (BamHI linearized)
pMCS5
pT7Blue
M13mp19
pEZ BAC (HpaI linearized)
pMiniT
pT7Blue-2
MBP
pF1A T7
pMiniT (linearized)
pT7Blue-3
pACYC177
pF1K T7
pMiniT 2.0
pT7CFE1-CFtag
pACYC184
pF25A ICE T7
pMiniT 2.0 (linearized)
pT7CFE1-CGFP-HA-His
pAN7-1
pF25K ICE T7
pMMB190
pT7CFE1-CGST-HA-His
pANT7_cGST
pF3A WG (BYDV)
pMMB206
pT7CFE1-CHA
pANT7_nHA
pF3K WG (BYDV)
pMMB207
pT7CFE1-CHis
pAUR316
pFC20A HaloTag T7 SP6
pMMB208
pT7CFE1-CMyc
pBC KS(+)
pFC20K HaloTag T7 SP6
pMMB66EH
pT7CFE1-NFtag
pBC KS(-)
pFC30A His6HaloTag T7
pMMB66HE
pT7CFE1-NHA
pBC SK(+)
pFC30K His6HaloTag T7
pMMB67EH
pT7CFE1-NHA-CHis
pBC SK(-)
pFC7A (HQ)
pMMB67HE
pT7CFE1-NHis
pBeloBAC11
pFC7K (HQ)
pNEB193
pT7CFE1-NHis-GST
pBluescript II KS(+)
pFN18A HaloTag T7
pNEB206A (linearized)
pT7CFE1-NHis-GST-CHA
pBluescript II KS(-)
pFN18K HaloTag T7
pOSIP-CH
pT7CFE1-NMyc
pBluescript II SK(+)
pFN19A HaloTag T7 SP6
pOSIP-CO
pTOP Blunt V2 (linearized)
pBluescript II SK(-)
pFN19K HaloTag T7 SP6
pOSIP-CT
pTrcHis A
pBluescript KS(+)
pFN29A His6HaloTag T7
pOSIP-KC
pTrcHis B
pBluescript KS(-)
pFN29K His6HaloTag T7
pOSIP-KH
pTrcHis C
pBluescript SK(+)
pFN2A (GST)
pOSIP-KL
pTrcHis2 A
pBluescript SK(-)
pFN2K (GST)
pOSIP-KO
pTrcHis2 B
pBR322
pFN6A (HQ)
pOSIP-KP
pTrcHis2 C
pBS KS(+)
pFN6K (HQ)
pOSIP-KT
pTV 118N
pBS KS(-)
pGEM-11Zf(+)
pOSIP-TH
pTWV228
pBS SK(+)
pGEM-11Zf(-)
pOSIP-TO
pTXB1
pBS SK(-)
pGEM-3Z
pOSIP-TT
pTYB21
pBS(+)
pGEM-3Zf(+)
pPCR-Script Amp SK(+)
pType IIs
pBS(-)
pGEM-3Zf(-)
pPCR-Script Cam SK(+)
pTZ19R
pBSK(+) Simple-Amp
pGEM-4Z
pPTR I
pTZ19U
pBSK(+) Simple-Kan
pGEM-5Zf(+)
pPTR II
pTZ57R
pcDNA3
pGEM-7Zf(+)
pRACE (linearized)
pUAST
pcDNA3.1(+)
pGEM-7Zf(-)
pRANGER-BTB-1
pUC118
pcDNA3.1(-)
pGEM-9Zf(-)
pRANGER-BTB-2
pUC119
pCold I
pGEM-T
pRANGER-BTB-3
pUC18
pCold II
pGEM-T (linearized)
pRANGER-BTB-5
pUC19
pCold III
pGEM-T Easy
pRHA-109
pUC19L
pCold IV
pGEM-T Easy (linearized)
pRHA-113
pUC57
pCold TF
pGGA
pRHA-67
pUC57-Kan
pCR-XL-2-TOPO
pGGAselect
pRSET A
pUC57-Simple
pCR-XL-2-TOPO (linearized)
PGK promoter
pRSET B
pUCmu
pDD
pGS-21a
pRSET C
pVLT31
pDNR-Dual
pH6HTC His6HaloTag T7
pSB1C3
pVLT33
pDNR-LIB
pH6HTN His6HaloTag T7
pSC101
pVLT35
pE-SUMOpro Amp
pHP45(omega)
pSEAP2-Basic
pYES1L
pE-SUMOpro Kan
pHP45(omega)-Km
pSF-Core
pZErO-2
pE-SUMOpro3 Amp
pHP45(omega)-Tc
pSMART BAC v2.0
SNAP-tag
pE-SUMOpro3 Kan
pHSG298
pSMART BAC v2.0 (BamHI linearized)
SP6 promoter
pE-SUMOstar Amp
pHSG299
pSMART BAC v2.0 (EcoRI linearized)
SV40 promoter
pE-SUMOstar Kan
pHSG396
pSMART BAC v2.0 (HindIII linearized)
T3 promoter
pEASY-Blunt (linearized)
pHSG398
pSMART FOS v2.0 (linearized)
T5 promoter
pEASY-Blunt E1 (linearized)
pHY300PLK
pSMART HCAmp
T7 promoter
pEASY-Blunt E2 (linearized)
pICOz
pSMART HCAmp (linearized)
tac promoter
pEASY-Blunt Simple (linearized)
pInvRecA
pSMART HCKan
trc promoter
pEASY-Blunt Zero (linearized)
pJAZZ-OC
pSMART HCKan (linearized)
phiX174
pEASY-Blunt3 (linearized)
pJAZZ-OK

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