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pAxCAwtit2 (#RDB05213)

Dual cassette vector to generate recombinant adenovirus containing CA promoter

Clone info. A dual cassette for constructing recombinant adenovirus containing CAG promoter. Improved version of pAxCAwtit. 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 pAxCAwtit2 cosmid is a derivative of pAxcwit2 (RDB no. 5212) and includes a CAG cassette [Niwa et al., Gene 108, 193-99, 1991], harboring a unique SwaI site as the cloning site for the gene of interest, at the E1 region. The pAxCAwtit2 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)
Selectable markers Amp^r
Gene/insert name AdV_5 - Genomic DNA
Depositor|Developer Saito, Izumu |
 
Remarks, protocol and/or map (pdf) RDB05213.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.
Ordering forms
Order form [Credit Card Payment] [Bank Transfer Payment] [Example of order form ]
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.
提供案内 (日本国内) [open/close]

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.)
RDB05213 pAxCAwtit2 DNA solution JPY 9,460 (not-for-profit academic purpose)
plus cost of shipping containers, dry ice (if required) and shipping charge

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 pAxCAwtit2 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB05213).

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 RDB13981_B1Lrp1-1.pdf check

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

Primer: CAG_R1 (Pr0584)
Region: CAG pro,beta-globin pA
Sequence file: RDB13981_B1Lra.seq check
>05213_13981_B1Lr_1_CAG_R1_B01_02_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNCT CNNGCGGGCG CGGGGCGAAG CGGTGCGGCG CCGGCAGGAA
   61 GGAAATGGGC GGGGAGGGCC TTCGTGCGTC GCCGCGCCGC CGTCCCCTTC TCCCTCTCCA
  121 GCCTCGGGGC TGTCCGCGGG GGGACGGCTG CCTTCGGGGG GGACGGGGCA GGGCGGGGTT
  181 CGGCTTCTGG CGTGTGACCG GCGGCTCTAG AGCCTCTGCT AACCATGTTC ATGCCTTCTT
  241 CTTTTTCCTA CAGCTCCTGG GCAACGTGCT GGTTATTGTG CTGTCTCATC ATTTTGGCAA
  301 AGAATTGATT TATCGATTTA AATTATAAAC TAGTCTAGAA TCGATAATCA ATTCACTCCT
  361 CAGGTGCAGG CTGCCTATCA GAAGGTGGTG GCTGGTGTGG CCAATGCCCT GGCTCACAAA
  421 TACCACTGAG ATCTTTTTCC CTCTGCCAAA AATTATGGGG ACATCATGAA GCCCCTTGAG
  481 CATCTGACTT CTGGCTAATA AAGGAAATTT ATTTTCATTG CAATAGTGTG TTGGAATTTT
  541 TTGTGTCTCT CACTCGGAAG GACATATGGG AGGGCAAATC ATTTAAAACA TCAGAATGAG
  601 TATTTGGTTT AGAGTTTGGC AACATATGCC NNTATGCTGG CTGNNNTGAA CAAAGGNTGG
  661 CTATNAAGAN GNCNTCAGTA TATGNANNNN CCCCCNGCTG TNNNNTNCNN TANNNNNTAN
721 AAAANNNNNN NNNN
//
Primer: Amp_R (Pr0008)
Region: AmpR proITR.1,Ad5 psi
Sequence file: RDB13981_B1Lrb.seq check
>05213_13981_B1Lr_1_Amp_R_C01_03_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNTTTATCN GGGTTATTGT CTCNTGAGNN NNNNCATATT
   61 TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC
  121 ACCTGACGTC TAAGAAACCA TTATTATCAT GACATTAACC TATAAAAATA GGCGTATCAC
  181 GAGGCCCTTT CGTCTTCAAG AATTCCGGAT CGATCCGCAT GTTCGATTTC GATTTAATTA
  241 ATTCGAACAT CATCAATAAT ATACCTTATT TTGGATTGAA GCCAATATGA TAATGAGGGG
  301 GTGGAGTTTG TGACGTGGCG CGGGGCGTGG GAACGGGGCG GGTGACGTAG TAGTGTGGCG
  361 GAAGTGTGAT GTTGCAAGTG TGGCGGAACA CATGTAAGCG ACGGATGTGG CAAAAGTGAC
  421 GTTTTTGGTG TGCGCCGGTG TACACAGGAA GTGACAATTT TCGCGCGGTT TTAGGCGGAT
  481 GTTGTAGTAA ATTTGGGCGT AACCGAGTAA GATTTGGCCA TTTTCGCGGG AAAACTGAAT
  541 AAGAGGAAGT GAAATCTGAA TAATTTTGTG TTACTCATAG CGCGTAATAT TTGTCTAGGG
  601 CCGCGGGGNN TTTGANCGTT TACGTGGAGN CTCGCCCANG TGNTTTTCTC NNGTGTTTTC
  661 CGCGTNNCGG GNCAAANTTG GCGTTTTNNT ANTATAGTCN GCATCNNTTN ANCNNCNNNN
721 NCNNNNNNGG NNNNNNGNNN NNNNNNN
//
Primer: AxIT_R (PrPr347)
Region: ITR.2
Sequence file: RDB13981_B1Lrc.seq check
>05213_13981_B1Lr_1_AxIT_R_D01_04_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNAAAC CNACAACTTC CTCAAATCGT CACTTCCGTT
   61 TTCCCACGTT ACGTCACTTC CCATTTTAAG AAAACTACAA TTCCCAACAC ATACAAGTTA
  121 CTCCGCCCTA AAACCTACGT CACCCGCCCC GTTCCCACGC CCCGCGCCAC GTCACAAACT
  181 CCACCCCCTC ATTATCATAT TGGCTTCAAT CCAAAATAAG GTATATTATT GATGATGTTC
  241 GAATTAATTA AATCGAAATC GAACATGCGG ATCCTCTAGA GTCAACAGCA GAAACATACA
  301 AGCTGTCAGC TTTGCACAAG GGCCCAACAC CCTGCTCATC AAGAAGCACT GTGGTTGCTG
  361 TGTTAGTAAT GTGCAAAACA GGAGGCACAT TTTCCCCACC TGTGTAGGTT CCAAAATATC
  421 TAGTGTTTTC ATTTTTACTT GGATCAGGAA CCCAGCACTC CACTGGATAA GCATTATCCT
  481 TATCCAAAAC AGCCTTGTGG TCAGTGTTCA TCTGCTGACT GTCGACCTGC AGGCATGCAA
  541 GCTTTAATGC GGTAGTTTAT CACAGTTAAA TTGCTAACGC AGTCAGGCAC CGTGTATTAA
  601 ATCTAACAAT GCGCTCATCG TCATCCTCGG CACCGTCACC CTGNANGCTG TANGCATANG
  661 CTNNNNTATG CCNGNACTGN CGGGCCTCTT GNGGGANATC GTCCATTNCN NANNNCATCN
721 NNNGNCACNN NNNNNNGNNN CNANCGNNNN NNNNNN
//

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 Kurihara, C., An easy method for preparation of Cre-loxP regulated fluorescent adenoviral expression vectors and its application for direct reprogramming into hepatocytes. Biotechnol. Rep. (Amst.) 12: 26-32 (2016). PMID 28352551. [link to RRC of NBRP]