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pAxCALNLwtit2 (#RDB05214)

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

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 transforming E. coli host)
Size of vector backbone 11 kb
Selectable markers Amp^r
Gene/insert name AdV_5 - Genomic DNA
Depositor|Developer Saito, Izumu |
 
Remarks, protocol and/or map (pdf) RDB05214.pdf

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
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MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ]
Please visit Information of Request for Distribution.[link] For for-profit-research purpose, please contact us. 
Terms and conditions for distribution 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:
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.
Remarks Remember that you will be working with samples containing infectious virus.
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必要書類
提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
手続きの概要は、「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。
MTAに書く使用条件 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))。
MTAに書く付加的使用条件：
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。
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.

How to cite this biological resource

Materials & Methods section:

The pAxCALNLwtit2 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB05214).

Reference section:

Fukuda, H., Terashima, M., Koshikawa, M., Kanegae, Y., Saito, I., Possible mechanism of adenovirus generation from a cloned viral genome tagged with nucleotides at its ends. Microbiol. Immunol., 50, 643-654 (2006). PMID 16924150. [link to RRC of NBRP]
Terashima, M., Kondo, S., Kanegae, Y., Saito, I., Exp. Med., 21, 931-936 (2003).

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

QC test results

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. [link to RRC of NBRP]
original Terashima, M., Exp. Med., 21, 931-936 (2003).