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CSII-CMV-RfA · PDF file CSII-CMV-RfA 10265 bp MluI 228 NheI 3165 Eco47III 3168 AgeI 3174 XhoI 3180 NotI 3318 BamHI 3389 EcoRI 3639 MluI 4086 BamHI 4092 PstI 4761 PstI 4905 NotI 4910

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Text of CSII-CMV-RfA · PDF file CSII-CMV-RfA 10265 bp MluI 228 NheI 3165 Eco47III 3168 AgeI 3174 XhoI...

  • CSII-CMV-RfA

    10265 bp

    MluI 228

    NheI 3165 Eco47III 3168 AgeI 3174 XhoI 3180

    NotI 3318 BamHI 3389

    EcoRI 3639 MluI 4086 BamHI 4092

    PstI 4761

    PstI 4905 NotI 4910

    BamHI 4918 XbaI 4924

    HpaI 4930

    ApaI 6247

    CMV RU5 5' SD

    RRE

    3' SA cPPT+CTS

    CMV

    RfA

    attR1

    Cm

    ccdBattR2PREdel U3 RU5

    BGH 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-RfA Plasmid size: 10262 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 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). PRE: Woodchuck hepatitis virus posttranscriptional regulatory element del U3: Deletion of enhancer and promoter sequences in the U3 region 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