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pmRi-ZsGreen1

Vector for doxycycline-controlled co-expression of a microRNA with ZsGreen1.

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pmRi-ZsGreen1 Sequence and MappmRi-ZsGreen1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 EcoO109I (3286) AatII (3232) ZraI (3230) SspI (3114) EarI (3105) XmnI (2909) ScaI (2790) PvuI (2680) FspI (2532) AseI (2482) BsaI (2371) AlwNI (1833) PspFI (1725) BbsI (3288) StuI (277) Eco53kI (292) SacI (294) EcoRI (323) BtgI - NcoI (333) BclI * (417) SgrAI (422) PflFI - Tth111I (569) NgoMIV (599) NaeI (601) KasI (638) NarI (639) SfoI (640) PluTI (642) BseRI (695) BmgBI (826) BfuAI - BspMI (846) BsrGI (885) BtgZI (988) BspEI (1028) BamHI (1040) PvuII (1054) MluI (1058) NheI (1064) BmtI (1068) EagI - NotI (1071) BspDI - ClaI (1079) HindIII (1084) EcoRV (1098) XbaI (1102) BsaBI * (1113) MfeI (1201) BsmI (1202) HpaI (1214) PsiI (1234) PciI (1417) DrdI (1525) BseYI (1721) pmRi-ZsGreen1 3301 bp
EcoO109I  (3286)
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.
AatII  (3232)
1 site
G A C G T C C T G C A G
ZraI  (3230)
1 site
G A C G T C C T G C A G
SspI  (3114)
1 site
A A T A T T T T A T A A
EarI  (3105)
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  (2909)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2790)
1 site
A G T A C T T C A T G A
PvuI  (2680)
1 site
C G A T C G G C T A G C
FspI  (2532)
1 site
T G C G C A A C G C G T
AseI  (2482)
1 site
A T T A A T T A A T T A
BsaI  (2371)
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.
AlwNI  (1833)
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.
PspFI  (1725)
1 site
C C C A G C G G G T C G
BbsI  (3288)
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.
StuI  (277)
1 site
A G G C C T T C C G G A
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
BtgI  (333)
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  (333)
1 site
C C A T G G G G T A C C
BclI  (417)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
SgrAI  (422)
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.
PflFI  (569)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (569)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
NgoMIV  (599)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (601)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
KasI  (638)
1 site
G G C G C C C C G C G G
NarI  (639)
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  (640)
1 site
G G C G C C C C G C G G
PluTI  (642)
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.
BseRI  (695)
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.
BmgBI  (826)
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.
BfuAI  (846)
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  (846)
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.
BsrGI  (885)
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.
BtgZI  (988)
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.
BspEI  (1028)
1 site
T C C G G A A G G C C T
BamHI  (1040)
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.
PvuII  (1054)
1 site
C A G C T G G T C G A C
MluI  (1058)
1 site
A C G C G T T G C G C A
NheI  (1064)
1 site
G C T A G C C G A T C G
BmtI  (1068)
1 site
G C T A G C C G A T C G
EagI  (1071)
1 site
C G G C C G G C C G G C
NotI  (1071)
1 site
G C G G C C G C C G C C G G C G
BspDI  (1079)
1 site
A T C G A T T A G C T A
ClaI  (1079)
1 site
A T C G A T T A G C T A
HindIII  (1084)
1 site
A A G C T T T T C G A A
EcoRV  (1098)
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  (1102)
1 site
T C T A G A A G A T C T
BsaBI  (1113)
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  (1201)
1 site
C A A T T G G T T A A C
BsmI  (1202)
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  (1214)
1 site
G T T A A C C A A T T G
PsiI  (1234)
1 site
T T A T A A A A T A T T
PciI  (1417)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (1525)
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.
BseYI  (1721)
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.
AmpR
2237 .. 3097  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2237 .. 3028  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2237 .. 3097  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3029 .. 3097  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2237 .. 3097  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ZsGreen1
335 .. 1027  =  693 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
335 .. 1027  =  693 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ori
1478 .. 2066  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1478 .. 2066  =  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
3098 .. 3202  =  105 bp
AmpR promoter
3098 .. 3202  =  105 bp
SV40 poly(A) signal
1215 .. 1296  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1215 .. 1296  =  82 bp
SV40 polyadenylation signal
MCS
1040 .. 1107  =  68 bp
multiple cloning site
MCS
1040 .. 1107  =  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
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