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Plasmid Files

pmOrange2

Vector for expressing mOrange2 in bacteria.

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pmOrange2.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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BspQI - SapI (3330) AflIII - PciI (3213) AlwNI (2804) BmrI (2285) BsaI (2259) BpmI (2256) PvuI (1956) TsoI (1927) ScaI (1844) XmnI (1725) HindIII (234) BfuAI - BspMI (239) SphI (244) BamHI (264) TspMI - XmaI (269) SmaI (271) Acc65I (273) AgeI (276) KpnI (277) MscI (312) FspAI (345) BlpI (427) BsaAI (512) BtgI - NcoI (715) BbsI (717) PflMI (721) KasI (867) NarI (868) SfoI (869) PluTI (871) BsgI (876) AleI (924) SgrAI (963) XcmI (968) BsrGI (982) NotI (994) ApoI - EcoRI (1008) SpeI (1072) BsiWI (1082) PspOMI (1087) ApaI (1091) PfoI (1145) NdeI (1283) BstAPI (1284) AflII (1340) ZraI (1404) AatII (1406) SspI (1520) pmOrange2 3338 bp
BspQI  (3330)
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  (3330)
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  (3213)
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  (3213)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (2804)
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.
BmrI  (2285)
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  (2259)
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  (2256)
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.
PvuI  (1956)
1 site
C G A T C G G C T A G C
TsoI  (1927)
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).
ScaI  (1844)
1 site
A G T A C T T C A T G A
XmnI  (1725)
1 site
G A A N N N N T T C C T T N N N N A A G
HindIII  (234)
1 site
A A G C T T T T C G A A
BfuAI  (239)
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  (239)
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.
SphI  (244)
1 site
G C A T G C C G T A C G
BamHI  (264)
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.
TspMI  (269)
1 site
C C C G G G G G G C C C
XmaI  (269)
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.
SmaI  (271)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (273)
1 site
G G T A C C C C A T G G
AgeI  (276)
1 site
A C C G G T T G G C C A
KpnI  (277)
1 site
G G T A C C C C A T G G
MscI  (312)
1 site
T G G C C A A C C G G T
FspAI  (345)
1 site
R T G C G C A Y Y A C G C G T R
BlpI  (427)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BsaAI  (512)
1 site
Y A C G T R R T G C A Y
BtgI  (715)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (715)
1 site
C C A T G G G G T A C C
BbsI  (717)
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.
PflMI  (721)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
KasI  (867)
1 site
G G C G C C C C G C G G
NarI  (868)
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  (869)
1 site
G G C G C C C C G C G G
PluTI  (871)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
BsgI  (876)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AleI  (924)
1 site
C A C N N N N G T G G T G N N N N C A C
SgrAI  (963)
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.
XcmI  (968)
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.
BsrGI  (982)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
NotI  (994)
1 site
G C G G C C G C C G C C G G C G
ApoI  (1008)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (1008)
1 site
G A A T T C C T T A A G
SpeI  (1072)
1 site
A C T A G T T G A T C A
BsiWI  (1082)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PspOMI  (1087)
1 site
G G G C C C C C C G G G
ApaI  (1091)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PfoI  (1145)
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.
NdeI  (1283)
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  (1284)
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.
AflII  (1340)
1 site
C T T A A G G A A T T C
ZraI  (1404)
1 site
G A C G T C C T G C A G
AatII  (1406)
1 site
G A C G T C C T G C A G
SspI  (1520)
1 site
A A T A T T T T A T A A
AmpR
1538 .. 2398  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1538 .. 1606  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1538 .. 2398  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1607 .. 2398  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1538 .. 2398  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
mOrange2
282 .. 992  =  711 bp
236 amino acids  =  26.8 kDa
Product: photostable monomeric orange derivative of DsRed fluorescent protein
mammalian codon-optimized
mOrange2
282 .. 992  =  711 bp
236 amino acids  =  26.8 kDa
Product: photostable monomeric orange derivative of DsRed fluorescent protein
mammalian codon-optimized
ori
2569 .. 3157  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2569 .. 3157  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
1433 .. 1537  =  105 bp
AmpR promoter
1433 .. 1537  =  105 bp
lac promoter
143 .. 173  =  31 bp
   Segment 1:  -35  
   143 .. 148  =  6 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
   Segment 2:  
   149 .. 166  =  18 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
   Segment 3:  -10  
   167 .. 173  =  7 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
181 .. 197  =  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
181 .. 197  =  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).
ATG
222 .. 224  =  3 bp
1 amino acid  =  155.2 Da
Product: start codon
ATG
222 .. 224  =  3 bp
1 amino acid  =  155.2 Da
Product: start codon
ORF:  1538 .. 2398  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  282 .. 992  =  711 bp
ORF:  236 amino acids  =  26.8 kDa
ORF:  144 .. 488  =  345 bp
ORF:  114 amino acids  =  11.9 kDa
ORF:  2002 .. 2268  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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