MluI 228 NotI 1521 1 CS-CDF-EG-PRE-RfAcPPT+CTS EF-1 GFP PRE RfA Cm attR1 ccdB attR2 R U5 BGH pA f1...

CS-CDF-EG-PRE-RfA11470 bp

MluI 228

NotI 1521

EcoRI 2565AgeI 2660

ApaI 3179

AgeI 3752

BspEI 4482XhoI 4509

NotI 5482BamHI 5553

BspEI 5799EcoRI 5803

MluI 6250BamHI 6256

PstI 6925

BspEI 7258

ApaI 7452

CMV RU55' SD

RRE

3' SAcPPT+CTS

EF-1

GFP

PRE

RfA

attR1CmccdB

attR2R

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: CS-CDF-EG-PRE-RfA Plasmid size: 11470 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 EF-1: Human elongation factor 1 α subunit promoter GFP: Enhanced green fluorescent protein (EGFP) 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

MluI 228 NotI 1521 1 CS-CDF-EG-PRE-RfAcPPT+CTS EF-1 GFP PRE RfA Cm attR1 ccdB attR2 R U5 BGH pA f1...

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