pTRE-Cycle2

Vector for expressing mCherry while cycling the level of a second protein by alternating expression and rapid degradation.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsMammalian Expression VectorspTRE-Cycle2
EcoRI (3813) AgeI (3807) NdeI (3802) SacII (3798) ApaI (3793) EcoO109I (3790) PspOMI (3789) BglII (3783) MscI (3745) BsaAI (3545) PstI - SbfI (3424) PflMI (3339) BbvCI - Bpu10I (3234) SgrAI - XcmI (3090) BsrGI (3071) XbaI (3060) AatII (2868) ZraI (2866) SspI (2750) EarI (2741) XmnI (2545) ScaI (2426) PvuI (2316) FspI (2168) AseI (2118) BsaI (2007) PaeR7I - PspXI - XhoI (1) Acc65I (335) KpnI (339) BfuAI - BspMI - PaqCI (351) BsmBI - Esp3I (377) BamHI (676) MluI (694) NheI (700) BmtI (704) EagI - NotI (707) BspDI - ClaI (715) HindIII (720) SalI (726) AccI (727) EcoRV (734) PciI (1053) pTRE-Cycle2 3892 bp
EcoRI  (3813)
1 site		 G A A T T C C T T A A G
AgeI  (3807)
1 site		 A C C G G T T G G C C A
NdeI  (3802)
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.
SacII  (3798)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
ApaI  (3793)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
EcoO109I  (3790)
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.
PspOMI  (3789)
1 site		 G G G C C C C C C G G G
BglII  (3783)
1 site		 A G A T C T T C T A G A
MscI  (3745)
1 site		 T G G C C A A C C G G T
BsaAI  (3545)
1 site		 Y A C G T R R T G C A Y
PstI  (3424)
1 site		 C T G C A G G A C G T C
SbfI  (3424)
1 site		 C C T G C A G G G G A C G T C C
PflMI  (3339)
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  (3234)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (3234)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
SgrAI  (3090)
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  (3090)
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  (3071)
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.
XbaI  (3060)
1 site		 T C T A G A A G A T C T
AatII  (2868)
1 site		 G A C G T C C T G C A G
ZraI  (2866)
1 site		 G A C G T C C T G C A G
SspI  (2750)
1 site		 A A T A T T T T A T A A
EarI  (2741)
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  (2545)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (2426)
1 site		 A G T A C T T C A T G A
PvuI  (2316)
1 site		 C G A T C G G C T A G C
FspI  (2168)
1 site		 T G C G C A A C G C G T
AseI  (2118)
1 site		 A T T A A T T A A T T A
BsaI  (2007)
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.
PaeR7I  (1)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1)
1 site		 C T C G A G G A G C T C
Acc65I  (335)
1 site		 G G T A C C C C A T G G
KpnI  (339)
1 site		 G G T A C C C C A T G G
BfuAI  (351)
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  (351)
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.
PaqCI  (351)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
BsmBI  (377)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (377)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
BamHI  (676)
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  (694)
1 site		 A C G C G T T G C G C A
NheI  (700)
1 site		 G C T A G C C G A T C G
BmtI  (704)
1 site		 G C T A G C C G A T C G
EagI  (707)
1 site		 C G G C C G G C C G G C
NotI  (707)
1 site		 G C G G C C G C C G C C G G C G
BspDI  (715)
1 site		 A T C G A T T A G C T A
ClaI  (715)
1 site		 A T C G A T T A G C T A
HindIII  (720)
1 site		 A A G C T T T T C G A A
SalI  (726)
1 site		 G T C G A C C A G C T G
AccI  (727)
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  (734)
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.
PciI  (1053)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AmpR
1873 .. 2733  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   1873 .. 2664  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1873 .. 2733  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   2665 .. 2733  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1873 .. 2733  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
mCherry
3066 .. 3776  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein
mammalian codon-optimized
mCherry
3066 .. 3776  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein
mammalian codon-optimized
ori
1114 .. 1702  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1114 .. 1702  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
bidirectional TRE promoter
3823 .. 318  =  388 bp
Tet-responsive bidirectional promoter PTight-BI, consisting of seven tet operator sequences flanked on each side by the minimal CMV promoter
bidirectional TRE promoter
3823 .. 318  =  388 bp
Tet-responsive bidirectional promoter PTight-BI, consisting of seven tet operator sequences flanked on each side by the minimal CMV promoter
DD
352 .. 675  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 1:  
   352 .. 354  =  3 bp
   1 amino acid  =  149.2 Da
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
352 .. 675  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 2:  
   355 .. 675  =  321 bp
   107 amino acids  =  11.8 kDa
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
352 .. 675  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
AmpR promoter
2734 .. 2838  =  105 bp
AmpR promoter
2734 .. 2838  =  105 bp
SV40 poly(A) signal
851 .. 932  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
851 .. 932  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2871 .. 2952  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2871 .. 2952  =  82 bp
SV40 polyadenylation signal
MCS
676 .. 737  =  62 bp
multiple cloning site
MCS
676 .. 737  =  62 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 .. 828  =  759 bp
ORF:  252 amino acids  =  28.2 kDa
ORF:  2003 .. 2269  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3804 .. 202  =  291 bp
ORF:  96 amino acids  =  11.0 kDa
ORF:  1873 .. 2733  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  3066 .. 3803  =  738 bp
ORF:  245 amino acids  =  27.7 kDa
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Individual Sequences & Maps

p3xFLAG-CMV-10
pcDNA6 myc-His A
pCMV6-AC-YFP
pHet-Act1-2
p3xFLAG-CMV-13
pcDNA6 myc-His B
pCMV6-AN-3DDK
pHet-Mem1
p3xFLAG-CMV-14
pcDNA6 myc-His C
pCMV6-AN-3HA
pHet-Nuc1
p3xFLAG-CMV-7
pcDNA6 TR
pCMV6-AN-DDK
pHom-1
p3xFLAG-CMV-7.1
pcDNA6 V5-His A
pCMV6-AN-FC
pHom-Mem1
p3xFLAG-CMV-8
pcDNA6 V5-His B
pCMV6-AN-FC-S
pHTC HaloTag CMV-neo
p3xFLAG-CMV-9
pcDNA6 V5-His C
pCMV6-AN-FP602
pHTN HaloTag CMV-neo
p3xFLAG-Myc-CMV-23
pCEP4
pCMV6-AN-FP635
pIRES
p3xFLAG-Myc-CMV-24
pCI
pCMV6-AN-GFP
pIRES2 DsRed-Express2
p3xFLAG-Myc-CMV-25
pCI-neo
pCMV6-AN-GFP-AC-His
pIRES2-AcGFP1
p3xFLAG-Myc-CMV-26
pCMV SPORT6
pCMV6-AN-HA
pIRES2-EGFP
pALTER-MAX
pCMV SPORT6.1
pCMV6-AN-His
pIRES2-ZsGreen1
pBApo-CMV
pCMV-(DYKDDDDK)-C
pCMV6-AN-His-DDK
pIRESbleo3
pBApo-CMV Neo
pCMV-(DYKDDDDK)-N
pCMV6-AN-His-HA
pIREShyg3
pBApo-CMV Pur
pCMV-3Tag-1A
pCMV6-AN-His-Myc
pIRESneo3
pBApo-EF1(alpha) Neo
pCMV-3Tag-1B
pCMV6-AN-mBFP
pIRESpuro3
pBApo-EF1(alpha) Pur
pCMV-3Tag-1C
pCMV6-AN-mCFP
pM-secSUMOstar
pBI-CMV1
pCMV-3Tag-2A
pCMV6-AN-mGFP
pM-SUMOstar
pBI-CMV2
pCMV-3Tag-2B
pCMV6-AN-mKate
pME-HA (linearized)
pBI-CMV3
pCMV-3Tag-2C
pCMV6-AN-mRFP
pmRi-mCherry
pBI-CMV4
pCMV-3Tag-3A
pCMV6-AN-Myc
pmRi-ZsGreen1
pBK-CMV
pCMV-3Tag-3B
pCMV6-AN-Myc-DDK
pNTAP-A
pBudCE4.1
pCMV-3Tag-3C
pCMV6-AN-mYFP
pNTAP-B
pcDNA3
pCMV-3Tag-4A
pCMV6-AN-RFP
pNTAP-C
pcDNA3.1+ C-DYK
pCMV-3Tag-4B
pCMV6-AN-YFP
pOG44
pcDNA3.1+ C-HA
pCMV-3Tag-4C
pCMV6-Entry
pPTuner
pcDNA3.1+ C-His
pCMV-3Tag-6
pCMV6-Entry-Cre
pPTuner IRES2
pcDNA3.1+ C-Myc
pCMV-3Tag-7
pCMV6-Entry2
pPTunerC
pcDNA3.1+ N-GST(TEV)
pCMV-3Tag-8
pCMV6-XL5-DDK-His
pREP4
pcDNA3.1+ N-GST(Thrombin)
pCMV-3Tag-9
pCS6-Empty
prHom-1
pcDNA3.1+ N-HA
pCMV-DDK-C
pCTAP-A
prHom-Nuc1
pcDNA3.1+ N-His
pCMV-DDK-N
pCTAP-B
prHom-Sec1
pcDNA3.1+ N-Myc
pCMV-HA
pCTAP-C
pSecTag A
pcDNA-DEST40
pCMV-HA-C
pDEST26
pSecTag B
pcDNA-DEST47
pCMV-HA-N
pDEST27
pSecTag C
pcDNA-DEST53
pCMV-LacZ
pDisplay
pSecTag2 A
pcDNA3.1(+)
pCMV-MIR
pEF-DEST51
pSecTag2 B
pcDNA3.1(-)
pCMV-Myc
pEF1alpha-IRES
pSecTag2 C
pcDNA3.1 CT-GFP
pCMV-Myc-C
pEF1alpha-IRES-AcGFP1
pSI
pcDNA3.1 His A
pCMV-Myc-N
pEF1alpha-IRES-DsRed-Express2
pSingle-tTs-shRNA
pcDNA3.1 His B
pCMV-Script
pEF1alpha-IRES-ZsGreen1
pT-REx-DEST30
pcDNA3.1 His C
pCMV-Script EX
pEF1alpha-Tet3G
pT-REx-DEST31
pcDNA3.1 Hygro(+)
pCMV-Tag 1
pEF5 FRT V5-DEST
pTandem-1
pcDNA3.1 Hygro(-)
pCMV-Tag 2A
pEF6 V5-His A
pTCN
pcDNA3.1 myc-His A
pCMV-Tag 2B
pEF6 V5-His B
pTCP
pcDNA3.1 myc-His B
pCMV-Tag 2C
pEF6 V5-His C
pTet-DualOFF
pcDNA3.1 myc-His C
pCMV-Tag 3A
pEntry-rtTA
pTet-DualON
pcDNA3.1 myc-His(-) A
pCMV-Tag 3B
pF4A CMV
pTet-Off
pcDNA3.1 myc-His(-) B
pCMV-Tag 3C
pF4K CMV
pTet-Off-Advanced
pcDNA3.1 myc-His(-) C
pCMV-Tag 4A
pF5A CMV-neo
pTet-On
pcDNA3.1 NT-GFP
pCMV-Tag 4B
pF5K CMV-neo
pTet-On-Advanced
pcDNA3.1 nV5-DEST
pCMV-Tag 4C
pFC14A HaloTag CMV
pTet-tTS
pcDNA3.1 V5-His A
pCMV-Tag 5A
pFC14K HaloTag CMV
pTetOne
pcDNA3.1 V5-His B
pCMV-Tag 5B
pFC15A HaloTag CMVd1
pTK-neo
pcDNA3.1 V5-His C
pCMV-Tag 5C
pFC15K HaloTag CMVd1
pTracer-CMV
pcDNA3.1 Zeo(+)
pCMV-Tet3G
pFC16A HaloTag CMVd2
pTracer-CMV Bsd
pcDNA3.1 Zeo(-)
pCMV6-A-BSD
pFC16K HaloTag CMVd2
pTracer-CMV2
pcDNA3.2 V5-DEST
pCMV6-A-EM7-BSD
pFC17A HaloTag CMVd3
pTracer-EF V5-His A
pcDNA4 His A
pCMV6-A-GFP
pFC17K HaloTag CMVd3
pTracer-EF V5-His B
pcDNA4 His B
pCMV6-A-Hygro
pFC27A HaloTag CMV-neo
pTracer-EF V5-His C
pcDNA4 His C
pCMV6-A-Puro
pFC27K HaloTag CMV-neo
pTracer-SV40
pcDNA4 HisMax A
pCMV6-AC
pFLAG-CMV-1
pTRE-Cycle1
pcDNA4 HisMax B
pCMV6-AC-3DDK
pFLAG-CMV-2
pTRE-Cycle2
pcDNA4 HisMax C
pCMV6-AC-3HA
pFLAG-CMV-3
pTRE-Cycle3
pcDNA4 myc-His A
pCMV6-AC-DDK
pFLAG-CMV-4
pTRE-Dual1
pcDNA4 myc-His B
pCMV6-AC-DDK-His
pFLAG-CMV-5.1
pTRE-Dual2
pcDNA4 myc-His C
pCMV6-AC-FC
pFLAG-Myc-CMV-19
pTRE-Tight
pcDNA4 TO
pCMV6-AC-FC-S
pFLAG-Myc-CMV-20
pTRE-Tight-BI
pcDNA4 TO myc-His A
pCMV6-AC-FP602
pFLAG-Myc-CMV-21
pTRE-Tight-BI-AcGFP1
pcDNA4 TO myc-His B
pCMV6-AC-FP635
pFLAG-Myc-CMV-22
pTRE-Tight-BI-DsRed-Express
pcDNA4 TO myc-His C
pCMV6-AC-GFP
pFN21A HaloTag CMV
pTRE-Tight-BI-ZsGreen1
pcDNA4 V5-His A
pCMV6-AC-GFP-rtTA
pFN21K HaloTag CMV
pTRE2
pcDNA4 V5-His B
pCMV6-AC-HA
pFN22A HaloTag CMVd1
pTRE2hyg
pcDNA4 V5-His C
pCMV6-AC-HA-His
pFN22K HaloTag CMVd1
pTRE2pur
pcDNA5 FRT
pCMV6-AC-His
pFN23A HaloTag CMVd2
pTRE3G
pcDNA5 FRT TO
pCMV6-AC-IRES-GFP
pFN23K HaloTag CMVd2
pTRE3G-BI
pcDNA5 TO
pCMV6-AC-IRES-GFP-Puro
pFN24A HaloTag CMVd3
pTRE3G-BI-Het1
pcDNA6.2 C-EmGFP-DEST
pCMV6-AC-mBFP
pFN24K HaloTag CMVd3
pTRE3G-BI-mCherry
pcDNA6.2 C-YFP-DEST
pCMV6-AC-mCFP
pFN28A HaloTag CMV-neo
pTRE3G-BI-ZsGreen1
pcDNA6.2 cGeneBLAzer-DEST
pCMV6-AC-mGFP
pFN28K HaloTag CMV-neo
pTRE3G-IRES
pcDNA6.2 cLumio-DEST
pCMV6-AC-mKate
pFRT lacZeo
pTRE3G-mCherry
pcDNA6.2 EmGFP-Bsd V5-DEST
pCMV6-AC-mRFP
pFRT lacZeo2
pTRE3G-ZsGreen1
pcDNA6.2 N-EmGFP-DEST
pCMV6-AC-Myc
pGen2.1
pVAX1
pcDNA6.2 N-YFP-DEST
pCMV6-AC-Myc-DDK
phCMV1
pX330
pcDNA6.2 nGeneBLAzer-DEST
pCMV6-AC-Myc-His
phCMV2
pZeoSV2(+)
pcDNA6.2 nLumio-DEST
pCMV6-AC-mYFP
phCMV3
pZeoSV2(-)
pcDNA6.2 V5-DEST
pCMV6-AC-RFP
pHet-1
pZFHD1-2
pcDNA6.2 V5-pL-DEST

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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