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pAxCALNLwtit2

Dual cassette vector to generate recombinant adenovirus containing loxP sites and CA promoter

Catalog number RDB05214
Resource name pAxCALNLwtit2
Clone info. A dual cassette of adenovirus vector for ON-OFF regulation. Improved version of pAxCALNLw. Csp45I and PacI can be used to generate recombinant adenovirus by transfection. Conversion to recombinant adenovirus was confirmed with HEK293 cell (Nov, 2005).
Comment The pAxCALNLwtit2 cosmid is a derivative of pAxcwit2 (RDB no. 5212) and includes a CALNL cassette [Kanegae et al., Nucleic Acids Res. 23, 3816-21, 1995; Kanegae et al., Gene 181, 207-12, 1996] at the E1 region. The pAxCALNLwtit2 cosmid, digested with Csp45I or PacI, can generate rAd upon transfection of HEK293 cells and can also be used to generate rAd by the COS-TPC method [Miyake et al., Proc. Natl. Acad. Sci. USA 93, 1320-24, 1996]. The resultant rAd expresses the gene of interest via the Cre-mediated control of gene expression by an expression cassette that provides the on/off-switching reporter unit and is composed of a CA promoter [Niwa et al., Gene 108, 193-99, 1991.], a loxP sequence, a stuffer sequence, and a second loxP sequence, which is followed by the SwaI cloning site and the G polyA sequence, namely, a CALNL cassette [Kanegae et al., Nucleic Acids Res. 23, 3816-21, 1995; Kanegae et al., Gene 181, 207-12, 1996].
Vector backbone charomid 9-11 (Cosmid, use packaging extracts for transformation)
Size of vector backbone 11 kb
Selectable markers Amp^r
Gene/insert name AdV_5 - Genomic DNA
Depositor|Developer Saito, Izumu |
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External Database
AdV_5 -

            Reference sequence
              

            Distribution information

            Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
            Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested (Terashima, M., Exp. Med., 21, 931-936(2003)).
            Additional terms and conditions for distribution Regarding resources containing CAG promoter: In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation for the CAG promoter (Niwa, H., Yamamura, K., Miyazaki, J., Gene 108 : 193-200, 1991) and an acknowledgment to Dr. Jun-ich Miyazaki of the Osaka University are requested.
            Ordering Please visit Information of Request for Distribution.[link] For for-profit-research purpose, please contact us. 
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            Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
            Remarks Remember that you will be working with samples containing infectious virus.
            提供案内 (日本国内) [open/close]

            提供条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する (Terashima, M., Exp. Med., 21, 931-936(2003))。
            付加的提供条件 CAGプロモータを含むリソースについて: 利用者は、研究成果の公表にあたってCAGプロモータの文献 (Niwa, H., Yamamura, K., Miyazaki, J., Gene 108 : 193-200, 1991)を引用し、大阪大学 宮崎純一博士への謝辞の表明を必要とする。
            提供依頼 手続きの詳細は、「提供申込みについて[link]」をご覧ください。営利目的利用についてはお問い合わせください。
            提供依頼書 [Word]
            提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]
            備考 このリソースはウイルス粒子産生用のため、取扱いに注意が必要です。

            Catalog # Resource name Shipping form Fee (non-profit org.)
            RDB05214 pAxCALNLwtit2 DNA solution

            Please review the QC test results indicated by check icon below as well as clone information before placing your order.

            References and tips

            Electronic file

            Remarks, protocol and/or map (pdf) RDB05214.pdf

            Original reference

            original Fukuda, H., Possible mechanism of adenovirus generation from a cloned viral genome tagged with nucleotides at its ends. Microbiol. Immunol., 50, 643-654 (2006). PMID 16924150.
            original Terashima, M., Exp. Med., 21, 931-936 (2003).

            Further references such as user reports and related articles (go to bottom)

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            Sequence information

            RIKEN BRC has sequenced portions of this material for quality test.
            Please review the QC test results indicated by check icon as well as clone information before placing your order.

            Test sheet RDB12784_A4Dsp1.pdf check

            Nucleotide sequence of a portion of this resource (if available).

            Primer: pCAG-F_V2
            Sequence file: RDB12784_A4Dsa_v2.seq check
            >05214_12784_A4Ds-pCAG-F_V2_A04_01.ab1
                1 GCACAGTCAA TCTGTGCGGA GCCGAATCTG GGAGGCGCCG CCGCACCCCC TCTAGCGGGC
               61 GCGGGGCGAA GCGGTGCGGC GCCGGCAGGA AGGAAATGGG CGGGGAGGGC CTTCGTGCGT
              121 CGCCGCGCCG CCGTCCCCTT CTCCCTCTCC AGCCTCGGGG CTGTCCGCGG GGGGACGGCT
              181 GCCTTCGGGG GGGACGGGGC AGGGCGGGGT TCGGCTTCTG GCGTGTGACC GGCGGCTCTA
              241 GAGCCTCTGC TAACCATGTT CATGCCTTCT TCTTTTTCCT ACAGCTCCTG GGCAACGTGC
              301 TGGTTATTGT GCTGTCTCAT CATTTTGGCA AAGAATTGAT TAATTCGAGC GAACGCGTAT
              361 AACTTCGTAT AGCATACATT ATACGAAGTT ATCTCGAGTC GGATTTGATC TGATCAAGAG
              421 ACAGGATGAG GATCGTTTCG CATGATTGAA CAAGATGGAT TGCACGCAGG TTCTCCGGCC
              481 GCTTGGGTGG AGAGGCTATT CGGCTATGAC TGGGCACAAC AGACAATCGG CTGCTCTGAT
              541 GCCGCCGTGT TCCGGCTGTC AGCGCAGGGG CGCCCGGTTC TTTTTGTCAA GACCGACCTG
              601 TCCGGTGCCC TGAATGAACT GCAGGACGAG GCAGCGCGGC TATCGTGGCT GGCCACGACG
              661 GGCGTTCCTT GCGCAGCTGT GCTCGACGTT GTCACTGAAG CGGGAAGGGA CTGGCTGCTA
              721 TTGGGCGAAG TGCCGGGGCA GGATCTCCTG TCATCTCACC TTGCTCCTGC CGAGAAAGTA
              781 TCCATCATGG CTGATGCAAT GCGGCGGCTG CATACGCTTG ATCCGGCTAC CTGCCCATTC
              841 GACCACCAAG CGAAACATCG CATCGAGCGA GCACGTACTC GGATGGAAGC CGGTCTTGTC
              901 GATCAGATGA TCTGGACGAA GAGCATCAGG GGCTCGCGCC AGCCGAACTG TTCGCCAGGC
              961 TCAAGCGCGC ATGCCGACGC GAGGATCTCG TCGTGACCCA TGGCGATGCC TGCTTGCCGA
             1021 TATTCATGGG TGGAAGATGG CCGCTATTCC TGGAGATCCA TCGACCTGTA TGGCCGGCCT
             1081 GGGATGTATG TCGGAACGGC ATATCATGGA CATTACCGAT TTGTCGATCC ACAGTCACTA
            1141 TAGCTGTGAG TAGAAGACTA TTGATAGCGA CGCA
            //
            Primer: Neo-C_F
            Sequence file: RDB12784_A4Dsb_v2.seq check
            >05214_12784_A4Ds-Neo-C_F_B04_04.ab1
                1 CGTCTGTAAT TGCTGAGAGC TTGGCGGCGA ATGGGCTGAC CGCTTCCTCG TGCTTTACGG
               61 TATCGCCGCT CCCGATTCGC AGCGCATCGC CTTCTATCGC CTTCTTGACG AGTTCTTCTG
              121 AGCGGGACTC TGGGGTTCGC GAAATGACCG ACCAAGCGAC GCCCAACCTG CCATCACGAG
              181 ATTTCGATTC CACCGCCGCC TTCTATGAAA GGTTGGGCTT CGGAATCGTT TTCCGGGACG
              241 CCGGCTGGAT GATCCTCCAG CGCGGGGATC TCATGCTGGA GTTCTTCGCC CACCCCATCG
              301 ATAACTTGTT TATTGCAGCT TATAATGGTT ACAAATAAAG CAATAGCATC ACAAATTTCA
              361 CAAATAAAGC ATTTTTTTCA CTGCATTCTA GTTGTGGTTT GTCCAAACTC ATCAATGTAT
              421 CTTATCATGT CTGGATCAAT CCGAACGCGT ATAACTTCGT ATAGCATACA TTATACGAAG
              481 TTATCTCGAG TCGCTCGGTA CGATTTCCAT CGATTTAAAT TATAAACTAG TCTAGAATCG
              541 ATGGGGATCA AATCAATTCA CTCCTCAGGT GCAGGCTGCC TATCAGAAGG TGGTGGCTGG
              601 TGTGGCCAAT GCCCTGGCTC ACAAATACCA CTGAGATCTT TTTCCCTCTG CCAAAAATTA
              661 TGGGGACATC ATGAAGCCCC TTGAGCATCT GACTTCTGGC TAATAAAGGA AATTTATTTT
              721 CATTGCAATA GTGTGTTGGA ATTTTTTGTG TCTCTCACTC GGAATGACAT ATGGGAGGGC
              781 AAATCATTTA AAACATCAGA ATGAGTATTT GGGTTTAGAG TTTGGCAACA TATGCCCATA
              841 TGCTGGCTGC CATGAACAAA GGTTGGCTAT AAAGAGGTCA TCAGTATATG AACAGCCCCT
              901 GCTGTCCATC GTATCATAGA AAAGGCTGGA CTGGAGTAAG ATTTTTTTTA ATTGGTTGGT
              961 AATTTTTCTA CATCCTAAAT TTCTTACTGT TTACTAGCGG ACTTTACTCC TCCTGACTAC
             1021 TCAGTCTAGC GTACACCTTT CGTAGATCCT GACTGAAGCC AGCTGATCAG AGTCTGGACT
            1081 A
            //
            Primer: Amp_R
            Sequence file: RDB12784_A4Dsc_v2.seq check
            >05214_12784_A4Ds-Amp_R_C04_07.ab1
                1 CTCGTTTCAT ATTATTGAGC ATTTATCAGG GTTATTGTCT CATGAGCGGA TACATATTTG
               61 AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC ATTTCCCCGA AAAGTGCCAC
              121 CTGACGTCTA AGAAACCATT ATTATCATGA CATTAACCTA TAAAAATAGG CGTATCACGA
              181 GGCCCTTTCG TCTTCAAGAA TTCCGGATCG ATCCGCATGT TCGATTTCGA TTTAATTAAT
              241 TCGAACATCA TCAATAATAT ACCTTATTTT GGATTGAAGC CAATATGATA ATGAGGGGGT
              301 GGAGTTTGTG ACGTGGCGCG GGGCGTGGGA ACGGGGCGGG TGACGTAGTA GTGTGGCGGA
              361 AGTGTGATGT TGCAAGTGTG GCGGAACACA TGTAAGCGAC GGATGTGGCA AAAGTGACGT
              421 TTTTGGTGTG CGCCGGTGTA CACAGGAAGT GACAATTTTC GCGCGGTTTT AGGCGGATGT
              481 TGTAGTAAAT TTGGGCGTAA CCGAGTAAGA TTTGGCCATT TTCGCGGGAA AACTGAATAA
              541 GAGGAAGTGA AATCTGAATA ATTTTGTGTT ACTCATAGCG CGTAATATTT GTCTAGGGCC
              601 GCGGGGACTT TGACCGTTTA CGTGGAGACT CGCCCAGGTG TTTTTCTCAG GTGTTTTCCG
              661 CGTTCCGGGT CAAAGTTGGC GTTTTATTAT TATAGTCAGC ATCGATTTAG CTTGGGCTGC
              721 AGGTCGAGGG ATCTCCATAA GAGAAGAGGG ACAGCTATGA CTGGGAGTAG TCAGGAGAGG
              781 AGGAAAAATC TGGCTAGTAA AACATGTAAG GAAAATTTTA GGGATGTTAA AGAAAAAAAT
              841 AACACAAAAC AAAAATATAA AAAAAATCTA ACCTCAAGTC AAGGCTTTTC TATGGAATAA
              901 GAATGGACAG CAAGGGGGCT GTTTCATATA CTGATGACCT CTTTATAGCC AACCCTTTGT
              961 TCATGGGCAG CCAGCATATG GGCATATGTT TGCCAAACTC TAAACCCAAA TACTCATTCT
             1021 GATGTTTTAA TGATTTGCCC TTCCCATATG TCCTTCCGAG TGAGAAGACA CAAAAATTCC
             1081 AAACACACTA ATTGCCAATG AAATAATTGC ATTTAATAGC CAGAAAGTTC AGATGCCTTC
            1141 ACAGAGGGGA CTCTCTC
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Fukuda, H., Possible mechanism of adenovirus generation from a cloned viral genome tagged with nucleotides at its ends. Microbiol. Immunol., 50, 643-654 (2006). PMID 16924150.
            original Terashima, M., Exp. Med., 21, 931-936 (2003).

            2022.05.18

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