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Flamindo/pcDNA3

Expression vector of yellow fluorescent biosensor for cAMP, Flamindo.

Catalog number RDB15247
Resource name Flamindo/pcDNA3
Clone info. Expression vector of yellow fluorescent cAMP indicator, Flamindo. A variant of YFP, Citrine was interrupted by cAMP-binding domain of mouse Epac1 (157-316aa). For the compositon, refer Fig.1 of Kitaguchi, T., Biochem. J. 450: 365-373, 2013.
Use Flamindo (R279E)/pcDNA3 (RDB15248) as a negative control.
Vector backbone pcDNA3 (plasmid)
Size of vector backbone 5.4 kb
Selectable markers Amp^r (E. coli), Neo^r (mammalian cell)
Gene/insert name mouse Epac1 cDNA Aequorea victoria GFP cDNA
Depositor Miyawaki, Atsushi |
 
Other clones in our bank mouse Rapgef3 (NCBI Gene 223864) |

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 1. The RECIPIENT should obtain written approval from the DEPOSITOR using the APPROVAL FORM prior to entering the AGREEMENT with the RIKEN BRC.
2. A RECIPIENT who belongs to a non-profit organization may use the BIOLOGICAL RESOURCE for an academic research that is non-commercial.
3. In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR or an acknowledgment to the DEPOSITOR are requested.
Kitaguchi T, Oya M, Wada Y, Tsuboi T, Miyawaki A. (2013) Extracellular Calcium influx activates Adenylate Cyclase 1 and potentiates Insulin secretion in MIN6 cells. Biochem. J., 450 (2): 365-373.
4. The RECIPIENT agrees to provide a copy of the publication to the DEPOSITOR, oral or written, reporting use of the MATERIAL.
5. The Recipient’s Scientist agrees not to transfer the MATERIAL to any third party including any resource centers without the prior written consent of DEPOSITOR.
Ordering Please visit Information of Request for Distribution. 
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Material Transfer Agreement (MTA) [Word]
Schedule A (GFP-A) [Link]
Approval form by depositor [Word]
Remarks
Please use Schedule A (GFP-A) additionally.
Please obtain written approval of the depositor.
Distribution information (domestic users) [open/close]


Catalog # Resource name Shipping form Fee (non-profit org.)
RDB15247 Flamindo/pcDNA3 DNA solution

Ordering Information [in Japanese] [in English]
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

Original reference

original Kitaguchi, T., Extracellular calcium influx activates adenylate cyclase 1 and potentiates insulin secretion in MIN6 cells. Biochem. J. 450 (2): 365-373 (2013). PMID 23282092.

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

Featured content

Featured content Dr. Miyawaki's resources (English text)
Featured content Sensor and Visualization (English text)

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 RDB15247_A7H9p1-2.pdf check

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

Primer: CMV_Forward
Region: CMV pro,insert5'
Sequence file: RDB15247_A7H9a.seq check
>D04498F5-2_A7H9_CMV-Forward_G08_20_ABI24.ab1
    1 TGTCACTCCG CCCATTGACG CAATGGGCGG TAGGCGTGTA CGGTGGGAGG TCTATATAAG
   61 CAGAGCTCTC TGGCTAACTA GAGAACCCAC TGCTTACTGG CTTATCGAAA TTAATACGAC
  121 TCACTATAGG GAGACCCAAG CTTATGGTGA GCAAGGGCGA GGAGCTGTTC ACCGGGGTGG
  181 TGCCCATCCT GGTCGAGCTG GACGGCGACG TAAACGGCCA CAAGTTCAGC GTGTCCGGCG
  241 AGGGCGAGGG CGATGCCACC TACGGCAAGC TGACCCTGAA GTTCATCTGC ACCACCGGCA
  301 AGCTGCCCGT GCCCTGGCCC ACCCTCGTGA CCACCTTCGG CTACGGCCTG ATGTGCTTCG
  361 CCCGCTACCC CGACCACATG AAGCAGCACG ACTTCTTCAA GTCCGCCATG CCCGAAGGCT
  421 ACGTCCAGGA GCGCACCATC TTCTTCAAGG ACGACGGCAA CTACAAGACC CGCGCCGAGG
  481 TGAAGTTCGA GGGCGACACC CTGGTGAACC GCATCGAGCT GAAGGGCATC GACTTCAAGG
  541 AGGACGGCAA CATCCTGGGG CACAAGCTGG AGTACAACCT CCGCGGGGAG GAGCTTGTTG
  601 AGGCTATGGC CCTCCTGTCC CAGCGAGGGC CTGATGCCCT ACTCACCGTG GCGCTCCGGA
  661 AGCCCCCAGG TCAGCGTACG GATGAAGAAC TGGACCTGAT CTTTGAAGAG CTGCTGCATA
  721 TCAAGGCGGT GGCACACCTT TCTAACTCGG TGAAGCGGGA ACTAGCTGCT GTTCTGCTCT
  781 TTGAACCACA CAGCAAGGCA GGAACTGTGT TGTTCAGCCA GGGGGACAAG GGTACCTCGT
  841 GGTACATTAT CTGGAAGGGA TCTGTCAATG TGGTGACCCA TGGCAAGGGG GCTGGTGACC
  901 ACGTTGCACG AGGGAGATGA CTTTGGACAG CTGGCTCTGG TGAACGACGC ACCTCGGGCA
  961 GCCACCATCA TCCTTCGAGA AAAATAACTG TCACTTTCTG CGTGTGGACA AGCAGGACTT
 1021 TCAACCGCAT CATCAAAGGA TGTGGAAAGC CAAAAACCCA TGAGACTGGT AAGCAGGATC
1081 ACCAGCCACA ACCGTCTATA TCAATTGACC GAACAGTCGA GGATCGGTCA TTCAAAATG
//
Primer: BGH_rev2
Region: bGHp(A),insert3'
Sequence file: RDB15247_A7H9b.seq check
>D04498F5-2_A7H9_BGH_rev2_H08_23_ABI24.ab1
    1 CTCAAGTAGT ATGCACGGGG GAGGGGCAAC AACAGATGGC TGGCAACTAG AAGGCACAGT
   61 CGAGGCTGAT CAGCGAGCTC TAGCATTTAG GTGACACTAT AGAATAGGGC CCTCTAGATG
  121 CATGCTCGAG TTACTTGTAC AGCTCGTCCA TGCCGAGAGT GATCCCGGCG GCGGTCACGA
  181 ACTCCAGCAG GACCATGTGA TCGCGCTTCT CGTTGGGGTC TTTGCTCAGG GCGGACTGGT
  241 AGCTCAGGTA GTGGTTGTCG GGCAGCAGCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT
  301 GGTAGTGGTC GGCGAGCTGC ACGCTGCCGT CCTCGATGTT GTGGCGGATC TTGAAGTTCA
  361 CCTTGATGCC GTTCTTCTGC TTGTCGGCCA TGATATAGAC GTTGTGGCTG TTGAATTCTT
  421 CTTCCAGTCT CATGGTTTTT GCTTCCACAT CCTTGATGAT GCGGTTGAAG TCCTGCTTGT
  481 CCACACGCAG AAAGTGACAG TTATTTTCTC GAAGGATGAT GGTGGCTGCC CGAGGTGCGT
  541 CGTTCACCAG AGCCAGCTGT CCAAAGTCAT CTCCCTCGTG CAACGTGGTC ACCAGCCCCT
  601 TGCCATGGGT CACCACATTG ACAGATCCCT TCCAGATAAT GTACCACGAG GTACCCTTGT
  661 CCCCCTGGCT GAACAACACA GTTCCTGCCT TGCTGTGTGG TTCAAAGAGC AGAACAGCAG
  721 CTAGTTCCCG CTTCACCGAG TTAGAAAGGT GTGCCACCGC CTTGATATGC AGCAGCTCTT
  781 CAAAGATCAG GTCCAGTTCT TCATCCGTAC GCTGACCTGG GGGCTTCCGG AGCGCCACGG
  841 TGAGTAGGGC ATCAGGCCCT CGCTGGGACA GGAGGGCCAT AGCCTCAACA AGCTCCTCCC
  901 CGCGGAGGTT GTACTCCAGC TTGTGCCCCA GGATGTTGCC GTCCTCCTTG AAGTCGATGC
  961 CCTTCAGCTC GATGCGGGTT CACCAGGGTG TCGCCCTCGA ACTTTCACCT CGGCGCGGGT
 1021 CTTGTTAGAT GCGTCGTCCT TGAGAGATGG TTGCGCCTCC TTGACGTAGC CTTTCGGCAT
1081 GGCCGAACTG CAGAAGGTCG TGCCTGCCTT TCAATGCTGG ATTCCCG
//
Primer: SV40pro_F
Region: SV40pro_ori,NeoR
Sequence file: RDB15247_A7H9c.seq check
>D04498F5-2_A7H9_SV40pro_F_A09_03_ABI24.ab1
    1 CCTGGTCCTG GGCATTCAGA GTAGTGAGGA GGCTTTTTTG GAGGCCTAGG CTTTTGCAAA
   61 AAGCTCCCGG GAGCTTGTAT ATCCATTTTC GGATCTGATC AAGAGACAGG ATGAGGATCG
  121 TTTCGCATGA TTGAACAAGA TGGATTGCAC GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG
  181 CTATTCGGCT ATGACTGGGC ACAACAGACA ATCGGCTGCT CTGATGCCGC CGTGTTCCGG
  241 CTGTCAGCGC AGGGGCGCCC GGTTCTTTTT GTCAAGACCG ACCTGTCCGG TGCCCTGAAT
  301 GAACTGCAGG ACGAGGCAGC GCGGCTATCG TGGCTGGCCA CGACGGGCGT TCCTTGCGCA
  361 GCTGTGCTCG ACGTTGTCAC TGAAGCGGGA AGGGACTGGC TGCTATTGGG CGAAGTGCCG
  421 GGGCAGGATC TCCTGTCATC TCACCTTGCT CCTGCCGAGA AAGTATCCAT CATGGCTGAT
  481 GCAATGCGGC GGCTGCATAC GCTTGATCCG GCTACCTGCC CATTCGACCA CCAAGCGAAA
  541 CATCGCATCG AGCGAGCACG TACTCGGATG GAAGCCGGTC TTGTCGATCA GGATGATCTG
  601 GACGAAGAGC ATCAGGGGCT CGCGCCAGCC GAACTGTTCG CCAGGCTCAA GGCGCGCATG
  661 CCCGACGGCG AGGATCTCGT CGTGACCCAT GGCGATGCCT GCTTGCCGAA TATCATGGTG
  721 GAAAATGGCC GCTTTTCTGG ATTCATCGAC TGTGGCCGGC TGGGTGTGGC GGACCGCTAT
  781 CAGGACATAG CGTTGGCTAC CCGTGATATT GCTGAAGAGC TTGGCGGCGA ATGGGCTGAC
  841 CGCTTCCTCG TGCTTTACGG TATCGCCGCT CCCGATTCGC AGCGCATCGC CTTCTATCGC
  901 CTTCTTGACG AGTTCTTCTG AGCGGGACTC TGGGGGTTCG AAATGACCGA CCAAGCGACG
  961 CCCAACCCTG CCCATCACGA GATTTCGATT TCCACCGCCG CCCTTCTATG AAAGGTTGGG
 1021 CTTTCGGAAT TCGCTTTCCA GGACGCCGGC TGGATGAATC TTCAAGCGCG GGATCTCAAT
1081 GGCTGAGTAT CGCCGACCCG ACCTGTTTAT TGGCAGGCCT TTAGTA
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Kitaguchi, T., Extracellular calcium influx activates adenylate cyclase 1 and potentiates insulin secretion in MIN6 cells. Biochem. J. 450 (2): 365-373 (2013). PMID 23282092.
reference Odaka, H., Genetically-encoded yellow fluorescent cAMP indicator with an expanded dynamic range for dual-color imaging. PLoS One 9 (6): e100252 (2014). PMID 24959857.

2019.12.09

GNP_filter3_RDBDEP_html_191130.pl