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pmRi-mCherry

Vector for doxycycline-controlled co-expression of a microRNA with mCherry.

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pmRi-mCherry.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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AatII (3247) ZraI (3245) SspI (3129) EarI (3120) XmnI (2924) ScaI (2805) PvuI (2695) FspI (2547) AseI (2497) BglI (2445) BsaI (2386) BmrI (2365) EcoO109I (3301) Eco53kI (292) SacI (294) EcoRI (323) MscI (365) BsaAI (565) PstI - SbfI (690) PflMI (774) BbvCI - Bpu10I (873) BsgI (929) SgrAI (1016) XcmI (1021) BsrGI (1035) BspEI (1043) BamHI (1055) MluI (1073) NheI (1079) BmtI (1083) EagI - NotI (1086) BspDI - ClaI (1094) HindIII (1099) SalI (1105) AccI (1106) EcoRV (1113) XbaI (1117) BsaBI * (1128) MfeI (1216) BsmI (1217) HpaI (1229) PsiI (1249) PciI (1432) NspI (1436) pmRi-mCherry 3316 bp
AatII  (3247)
1 site
G A C G T C C T G C A G
ZraI  (3245)
1 site
G A C G T C C T G C A G
SspI  (3129)
1 site
A A T A T T T T A T A A
EarI  (3120)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI recognition sequence.
Sticky ends from different EarI sites may not be compatible.
XmnI  (2924)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2805)
1 site
A G T A C T T C A T G A
PvuI  (2695)
1 site
C G A T C G G C T A G C
FspI  (2547)
1 site
T G C G C A A C G C G T
AseI  (2497)
1 site
A T T A A T T A A T T A
BglI  (2445)
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.
BsaI  (2386)
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.
BmrI  (2365)
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.
EcoO109I  (3301)
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.
Eco53kI  (292)
1 site
G A G C T C C T C G A G
SacI  (294)
1 site
G A G C T C C T C G A G
EcoRI  (323)
1 site
G A A T T C C T T A A G
MscI  (365)
1 site
T G G C C A A C C G G T
BsaAI  (565)
1 site
Y A C G T R R T G C A Y
PstI  (690)
1 site
C T G C A G G A C G T C
SbfI  (690)
1 site
C C T G C A G G G G A C G T C C
PflMI  (774)
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.
BbvCI  (873)
1 site
C C T C A G C G G A G T C G
Bpu10I  (873)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BsgI  (929)
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).
SgrAI  (1016)
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  (1021)
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  (1035)
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.
BspEI  (1043)
1 site
T C C G G A A G G C C T
BamHI  (1055)
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.
MluI  (1073)
1 site
A C G C G T T G C G C A
NheI  (1079)
1 site
G C T A G C C G A T C G
BmtI  (1083)
1 site
G C T A G C C G A T C G
EagI  (1086)
1 site
C G G C C G G C C G G C
NotI  (1086)
1 site
G C G G C C G C C G C C G G C G
BspDI  (1094)
1 site
A T C G A T T A G C T A
ClaI  (1094)
1 site
A T C G A T T A G C T A
HindIII  (1099)
1 site
A A G C T T T T C G A A
SalI  (1105)
1 site
G T C G A C C A G C T G
AccI  (1106)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
EcoRV  (1113)
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.
XbaI  (1117)
1 site
T C T A G A A G A T C T
BsaBI  (1128)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MfeI  (1216)
1 site
C A A T T G G T T A A C
BsmI  (1217)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
HpaI  (1229)
1 site
G T T A A C C A A T T G
PsiI  (1249)
1 site
T T A T A A A A T A T T
PciI  (1432)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (1436)
1 site
R C A T G Y Y G T A C R
AmpR
2252 .. 3112  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2252 .. 3043  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2252 .. 3112  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3044 .. 3112  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2252 .. 3112  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
mCherry
335 .. 1042  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein
mammalian codon-optimized
mCherry
335 .. 1042  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein
mammalian codon-optimized
ori
1493 .. 2081  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1493 .. 2081  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
tight TRE promoter
4 .. 318  =  315 bp
Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter
tight TRE promoter
4 .. 318  =  315 bp
Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter
AmpR promoter
3113 .. 3217  =  105 bp
AmpR promoter
3113 .. 3217  =  105 bp
SV40 poly(A) signal
1230 .. 1311  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1230 .. 1311  =  82 bp
SV40 polyadenylation signal
MCS
1055 .. 1122  =  68 bp
multiple cloning site
MCS
1055 .. 1122  =  68 bp
multiple cloning site
tet operator
12 .. 30  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
12 .. 30  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
48 .. 66  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
48 .. 66  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
83 .. 101  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
83 .. 101  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
119 .. 137  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
119 .. 137  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
155 .. 173  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
155 .. 173  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
190 .. 208  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
190 .. 208  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
226 .. 244  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
226 .. 244  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  70 .. 360  =  291 bp
ORF:  96 amino acids  =  11.1 kDa
ORF:  335 .. 1054  =  720 bp
ORF:  239 amino acids  =  27.0 kDa
ORF:  2382 .. 2648  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3267 .. 202  =  252 bp
ORF:  83 amino acids  =  9.8 kDa
ORF:  2252 .. 3112  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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