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pAxEFwtit2 (#RDB05215)

Dual cassette vector to generate recombinant adenovirus containing EF1 alpha promoter

Clone info. A dual cassette for constructing recombinant adenovirus containing EF1alpha promoter. Csp45I and PacI cab be used to generate recombinant adenovirus by transfection. Conversion to recombinant adenovirus was confirmed with HEK293 cell (Nov, 2005).
Comment The pAxEFwtit2 contains an expression cassette, consisting of the promoter of a gene for elongation factor 1alpha (EF-1alpha), a unique SwaI site and a polyadenylation signal from SV40 (S polyA), and was cloned into the SwaI site of pAxcwit2 (RDB 5212). The pAxEFwtit2 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].
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) RDB05215.pdf

Distribution information

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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)).
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))。
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.)
RDB05215 pAxEFwtit2 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 pAxEFwtit2 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB05215).

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 RDB19476_B1Jfp1-1.pdf check

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

Primer: EF1a-C_F (Pr0168)
Region: EF-1alpha pro,SV40 pA
Sequence file: RDB19476_B1Jfa.seq check
>05215_19476_B1Jf_3_EF1a-C_F_C02_08_ABI24.ab1
    1 NNNNNNNNNN NNNTTTTTNA GTTTGGNNCT TGGTTNNTTC TNNANGCCTC AGACAGTGGT
   61 TCAAAGTTTT TTTCTTCCAT TTCAGGTGTC GTGAGGAATT AATTCATCGA TTTAAATTAT
  121 AAACTAGTCT AGAATCGATG AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA
  181 ATAGCATCAC AAATTTCACA NNNNNNGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT
  241 CCAAACTCAT CAATGTATCT TATCATGTCT GGATCTCGAG GGCCCAAGCT GTTTAAACGA
  301 TGCTGACTAT AATAATAAAA CGCCAACTTT GACCCGGAAC GCGGAAAACA CCTGAGAAAA
  361 ACACCTGGGC GAGTCTCCAC GTAAACGGTC AAAGTCCCCG CGGCCCTAGA CAAATATTAC
  421 GCGCTATGAG TAACACAAAA TTATTCAGAT TTCACTTCCT CTTATTCAGT TTTCCCGCGA
  481 AAATGGCCAA ATCTTACTCG GTTACGCCCA AATTTACTAC AACATCCGCC TAAAACCGCG
  541 CGAAAATTGT CACTTCCTGT GTACACCGGC GCACACNNAA AACGTCACTT TTGCCACATC
  601 CGTCGCTTAC ATGTGTTCCG CCACNCTNGC AANNTCNCAC TTNCGCCNCA CTACTACGTC
  661 NCCCNNCCCN NTTNCCNCGC CCNNNGCCNN GNCACAAANT NNNNCCCCTC NNNANNNNNN
721 NNNN
//
Primer: pJB8-Forward (Pr0084)
Region: ITR.1,Ad5 psi,
Sequence file: RDB19476_B1Jfb.seq check
>05215_19476_B1Jf_3_pJB8-Forward_D02_11_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNTTC NANTTAATTA ATTCGAACNT CATCAATAAT ATACCTTATT
   61 TTGGATTGAA GCCAATATGA TAATGAGGGG GTGGAGTTTG TGACGTGGCG CGGGGCGTGG
  121 GAACGGGGCG GGTGACGTAG TAGTGTGGCG GAAGTGTGAT GTTGCAAGTG TGGCGGAACA
  181 CATGTAAGCG ACGGATGTGG CAAAAGTGAC GTTTTTGGTG TGCGCCGGTG TACACAGGAA
  241 GTGACAATTT TCGCGCGGTT TTAGGCGGAT GTTGTAGTAA ATTTGGGCGT AACCGAGTAA
  301 GATTTGGCCA TTTTCGCGGG AAAACTGAAT AAGAGGAAGT GAAATCTGAA TAATTTTGTG
  361 TTACTCATAG CGCGTAATAT TTGTCTAGGG CCGCGGGGAC TTTGACCGTT TACGTGGAGA
  421 CTCGCCCAGG TGTTTTTCTC ANGTGTTTTC CGCGTTCCGG GTCAAAGTTG GCGTTTTATT
  481 ATTATAGTCA GCATCGTTTA AACAGCTTGG GCCCTCGAGA TCCAGACATG ATAAGATACA
  541 TTGATGAGTT TGGACAAACC ACAACTAGAA TGCAGTGAAA AAAATGCTTT ATTTGTGAAA
  601 TTTGTGATGC TATTGCTTTA TTTGTAACCN TTATAAGCTG CNATAAACAA GTTCNTCGAT
  661 TCTAGANNNN NTNANAANTT AAATCNNTGA NNTNATNCCN CNCNANNCCT GANNNNNNNN
721 AAAANNNNNN NNNN
//
Primer: AxIT_R (PrPr347)
Region: ITR.2
Sequence file: RDB19476_B1Jfc.seq check
>05215_19476_B1Jf_3_AxIT_R_H03_24_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NACTTCCTCN AATCGTCACT TCCGTTTTCC CACGTTACGT
   61 CACTTCCCAT TTTAAGAAAA CTACAATTCC CAACACATAC AAGTTACTCC GCCCTAAAAC
  121 CTACGTCACC CGCCCCGTTC CCACGCCCCG CGCCACGTCA CAAACTCCAC CCCCTCATTA
  181 TCATATTGGC TTCAATCCAA AATAAGGTAT ATTATTGATG ATGTTCGAAT TAATTAAATC
  241 GAAATCGAAC ATGCGGATCC TCTAGAGTCA ACAGCAGAAA CATACAAGCT GTCAGCTTTG
  301 CACAAGGGCC CAACACCCTG CTCATCAAGA AGCACTGTGG TTGCTGTGTT AGTAATGTGC
  361 AAAACAGGAG GCACATTTTC CCCACCTGTG TAGGTTCCAA AATATCTAGT GTTTTCATTT
  421 TTACTTGGAT CAGGAACCCA GCACTCCACT GGATAAGCAT TATCCTTATC CAAAACAGCC
  481 TTGTGGTCAG TGTTCATCTG CTGACTGTCG ACCTGCAGGC ATGCAAGCTT TAATGCGGTA
  541 GTTTATCACA GTTAAATTGC TAACGCAGTC AGGCACCGTG TATNNAAATC TAACAATGCG
  601 CTCATCGTCA TCCTCGGCNN CGTCACCCTG GNNGCTGTAG GCATNNNCNT NNNTNNGCNN
  661 GTNCNGCCNG GNCTNNNGCG GGNNANCGNC CANTNCNANN GCNNCNCCNN NNNCNNNNGG
721 NNNGNTGNNA NCNNNNNNNG NNTNNNNGCN NTTNNNANNN NNNNNNN
//

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).
user_report Kise, S., Functional analysis of vitamin D receptor (VDR) using adenovirus vector. J. Steroid Biochem. Mol. Biol. 230: 106275 (2023). PMID 36854350. [link to RRC of NBRP]
user_report Araki, Y., Efficient recombinant production in mammalian cells using a novel IR/MAR gene amplification method. PLoS One, 7 (7): e41787 (2012). PMID 22844523. [link to RRC of NBRP]