pmRi-ZsGreen1

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

Sequence Author: Clontech (TaKaRa)

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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

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
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
2 segments
   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
2 segments
   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
ORF:  70 .. 1050  =  981 bp
ORF:  326 amino acids  =  38.0 kDa
ORF:  335 .. 1039  =  705 bp
ORF:  234 amino acids  =  26.4 kDa
ORF:  2367 .. 2633  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3252 .. 202  =  252 bp
ORF:  83 amino acids  =  9.8 kDa
ORF:  2237 .. 3097  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  712 .. 1005  =  294 bp
ORF:  97 amino acids  =  10.9 kDa
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