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Plasmid Files

pMCentr1

Plasmid with a Zeocin™ resistance marker and a TEV cleavage site, for creating a Gateway® entry vector by ligation-independent cloning (LIC).

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pMCentr1 Sequence and MappMCentr1.dna
Map and Sequence File   
Sequence Author:  Midwest Center for Structural Genomics
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 PciI (2109) DrdI (2007) BssS α I (1936) BciVI (1911) AlwNI (1700) AcuI - Eco57MI (1567) BsaAI - PmlI (1377) NcoI - StyI (1306) MscI (1305) BssHII - MauBI (1268) AatII (1266) ZraI (1264) SmaI (1218) TspMI - XmaI (1216) SgrAI (1190) BmgBI (1174) SexAI * (1140) BanI (1134) NspI (2113) BspQI - SapI (111) Bts α I (214) TsoI (257) AclI (364) BbsI (437) HpaI (501) AhdI (549) AflII (554) PspOMI (563) EcoO109I (564) ApaI - BanII (567) MslI (618) XcmI (692) SspI (694) EcoRV (823) BseRI (935) DraIII (958) EagI (973) BglI (1006) FseI (1036) pMCentr1 2115 bp
PciI  (2109)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (2007)
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.
BssSαI  (1936)
1 site
C A C G A G G T G C T C
BciVI  (1911)
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.
AlwNI  (1700)
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.
AcuI  (1567)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
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).
Eco57MI  (1567)
1 site
C T G R A G ( N ) 14 N N G A C Y T C ( N ) 14

Sticky ends from different Eco57MI sites may not be compatible.
After cleavage, Eco57MI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaAI  (1377)
1 site
Y A C G T R R T G C A Y
PmlI  (1377)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
NcoI  (1306)
1 site
C C A T G G G G T A C C
StyI  (1306)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MscI  (1305)
1 site
T G G C C A A C C G G T
BssHII  (1268)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
MauBI  (1268)
1 site
C G C G C G C G G C G C G C G C
AatII  (1266)
1 site
G A C G T C C T G C A G
ZraI  (1264)
1 site
G A C G T C C T G C A G
SmaI  (1218)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (1216)
1 site
C C C G G G G G G C C C
XmaI  (1216)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SgrAI  (1190)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
BmgBI  (1174)
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.
SexAI  (1140)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BanI  (1134)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
NspI  (2113)
1 site
R C A T G Y Y G T A C R
BspQI  (111)
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  (111)
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.
BtsαI  (214)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
TsoI  (257)
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).
AclI  (364)
1 site
A A C G T T T T G C A A
BbsI  (437)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
HpaI  (501)
1 site
G T T A A C C A A T T G
AhdI  (549)
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.
AflII  (554)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
PspOMI  (563)
1 site
G G G C C C C C C G G G
EcoO109I  (564)
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.
ApaI  (567)
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  (567)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
MslI  (618)
1 site
C A Y N N N N R T G G T R N N N N Y A C
XcmI  (692)
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.
SspI  (694)
1 site
A A T A T T T T A T A A
EcoRV  (823)
1 site
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.
BseRI  (935)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
DraIII  (958)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
EagI  (973)
1 site
C G G C C G G C C G G C
BglI  (1006)
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.
FseI  (1036)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
ori
1465 .. 2053  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1465 .. 2053  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
BleoR
935 .. 1309  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
BleoR
935 .. 1309  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
attL1
569 .. 668  =  100 bp
recombination site for the Gateway® LR reaction
attL1
569 .. 668  =  100 bp
recombination site for the Gateway® LR reaction
attL2
711 .. 810  =  100 bp
recombination site for the Gateway® LR reaction
attL2
711 .. 810  =  100 bp
recombination site for the Gateway® LR reaction
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
EM7 promoter
1328 .. 1375  =  48 bp
synthetic bacterial promoter
EM7 promoter
1328 .. 1375  =  48 bp
synthetic bacterial promoter
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
TEV site
671 .. 691  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV site
671 .. 691  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
T7 promoter
828 .. 846  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
828 .. 846  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
537 .. 553  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
537 .. 553  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
851 .. 867  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
851 .. 867  =  17 bp
common sequencing primer, one of multiple similar
variants
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