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pAxEFwtit2

Dual cassette vector to generate recombinant adenovirus containing EF1 alpha promoter

Catalog number RDB05215
Resource name pAxEFwtit2
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).
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 transformation)
Size of vector backbone 11 kb
Selectable markers Amp^r
Gene/insert name AdV_5 - Genomic DNA
Depositor Saito, Izumu |

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)).
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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB05215 pAxEFwtit2 DNA solution

Ordering Information [in Japanese] [in English]

References and tips

Electronic file

Electronic file PDF RDB05215.pdf from Depositor

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References

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).

User reports and related articles (go to bottom)


Sequence information

RIKEN BRC has sequenced portions of this material for quality test. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB05215_A3H9p1.pdf

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

Primer: EF1a-C_F RDB05215_A3H9a.seq
>05215_05215_A3H9_EF1a-C_F_D04_10.ab1
    1 GTACTCCAGC TCAGACAGTG GTTCAAGTTT TTTTCTTCCA TTTCAGGTGT CGTGAGGAAT
   61 TAATTCATCG ATTTAAATTA TAAACTAGTC TAGAATCGAT GAACTTGTTT ATTGCAGCTT
  121 ATAATGGTTA CAAATAAAGC AATAGCATCA CAAATTTCAC AAATAAAGCA TTTTTTTCAC
  181 TGCATTCTAG TTGTGGTTTG TCCAAACTCA TCAATGTATC TTATCATGTC TGGATCTCGA
  241 GGGCCCAAGC TGTTTAAACG ATGCTGACTA TAATAATAAA ACGCCAACTT TGACCCGGAA
  301 CGCGGAAAAC ACCTGAGAAA AACACCTGGG CGAGTCTCCA CGTAAACGGT CAAAGTCCCC
  361 GCGGCCCTAG ACAAATATTA CGCGCTATGA GTAACACAAA ATTATTCAGA TTTCACTTCC
  421 TCTTATTCAG TTTTCCCGCG AAAATGGCCA AATCTTACTC GGTTACGCCC AAATTTACTA
  481 CAACATCCGC CTAAAACCGC GCGAAAATTG TCACTTCCTG TGTACACCGG CGCACACCAA
  541 AAACGTCACT TTTGCCACAT CCGTCGCTTA CATGTGTTCC GCCACACTTG CAACATCACA
  601 CTTCCGCCAC ACTACTACGT CACCCGCCCC GTTCCCACGC CCCGCGCCAC GTCACAAACT
  661 CCACCCCCTC ATTATCATAT TGGCTTCAAT CCAAAATAAG GTATATTATT GATGATGTTC
  721 GAATTAATTA AATCGAAATC GAACATGCGG ATCGATCCGG AATTCTTGAA GACGAAAGGG
  781 CCTCGTGATA CGCCTATTTT TATAGGTTAA TGTCATGATA ATAATGGTTT CTTAGACGTC
  841 AGGTGGCACT TTTTCGGGGA AATGTGCGCG GAACCCCTAT TTGTTTATTT TTTCTAAATA
  901 CATTCAAATA TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTTC AATAATATTG
  961 AAAAAGGAAG AGTATGAGTA TTCAACATTT CCGTGTCGCC CTTTATTCCC TTTTTTTGCG
 1021 GCATTTTGCC CTTCCTGTTT TTTGCTCACC CAGAACGCTG GTTGAAAGGT AAAGATGGCT
 1081 GGAGGATCAG TTGGATGCAC GAAGTTGGGT AACATCGACT GGAATTCTTC AAACAAGGCG
1141 GGTTACA
//

Please visit Sequencing and PCR primers for primer information.


References

References

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).
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.

2018.12.10

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