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CSII-CMV-mRFP1-RfA 10671 bp MluI 228 NheI 3165 Eco47III 3168 AgeI 3174 BamHI 3187 PstI 3530 EcoRI 3872 NotI 3878 BamHI 3886 XbaI 3892 HpaI 3898 NotI 4877 BamHI 4948 BspEI 5194 EcoRI 5198 MluI 5645 BamHI 5651 PstI 6320 BspEI 6459 ApaI 6653 CMV R U5 5' SD RRE 3' SA cPPT+CTS CMV mRFP1 PRE RfA attR1 Cm ccdB attR2 R U5 BGH pA f1 ori SV40 pro & ori Zeo SV40 pA ColE1 ori Amp 1

CSII-CMV-mRFP1-RfA 10671 bp · 2014-09-29 · 10671 bp MluI 228 NheI 3165 Eco47III 3168 AgeI 3174 BamHI 3187 PstI 3530 EcoRI 3872 NotI 3878 BamHI 3886 XbaI 3892 HpaI 3898 NotI 4877

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Page 1: CSII-CMV-mRFP1-RfA 10671 bp · 2014-09-29 · 10671 bp MluI 228 NheI 3165 Eco47III 3168 AgeI 3174 BamHI 3187 PstI 3530 EcoRI 3872 NotI 3878 BamHI 3886 XbaI 3892 HpaI 3898 NotI 4877

CSII-CMV-mRFP1-RfA10671 bp

MluI 228

NheI 3165Eco47III 3168AgeI 3174BamHI 3187

PstI 3530

EcoRI 3872NotI 3878BamHI 3886XbaI 3892HpaI 3898

NotI 4877BamHI 4948

BspEI 5194EcoRI 5198

MluI 5645BamHI 5651

PstI 6320BspEI 6459

ApaI 6653

CMV RU55' SD

RRE

3' SAcPPT+CTS

CMV

mRFP1

PRE

RfA

attR1CmccdBattR2R

U5BGH pA

f1 ori

SV40 pro & ori

Zeo

SV40 pA

ColE1 ori

Amp 1

Miyoshi
テキストボックス
Plasmid name: CSII-CMV-mRFP1-RfA Plasmid size: 10671 bp Note CMV: Human cytomegalovirus immediate early promoter y: Packaging signal 5’SD: 5’ splicing donor site 3’SA: 3’ splicing acceptor site RRE: Rev responsive element cPPT: Central polypurine tract CTS: Central termination sequence mRFP1: Monomeric red fluorescent protein gene PRE: Woodchuck hepatitis virus posttranscriptional regulatory element RfA in the U3 region RfA: Gateway Reading Frame Cassette A containing Cm, ccdB, attR1, and attR2 Cm: Chloramphenicol resistance gene ccdB: The ccdB gene encodes a protein that interferes with E. coli DNA gyrase attR1, 2: Sites for site-specific recombination with a Gateway vector (Invitrogen). attR sites always recombine with attL sites in a reaction mediated by the LR Clonase (Invitrogen). BGH pA: Bovine growth hormone polyadenylation signal SV40 pro & ori: SV40 early promoter and origin Zeo: Zeocin resistance gene SV40 pA: SV40 polyadenylation signal Amp: Ampicillin resistance gene *Use the DB3.1 or ccdB Survival 2 T1R E. coli strain for propagation of plasmids containing the ccdB gene.
Miyoshi
テキストボックス
y