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YC3.60/pcDNA3

Yellow cameleon calcium ion sensor clone YC3.60.

Catalog number RDB15135
Resource name YC3.60/pcDNA3
Clone info. Calcium ion sensor clone YC3.60 Yellow cameleon having E104Q substitution in the 3rd Ca2+ binding site of CaM and the circular permutated cp173Venus. Mammalian expression plasmid.
Known differences (at least): C to T at 291nt (97aa, Arg, synonym), T to C at 438nt (146aa, Tyr, synonym), A to C at 493nt (165aa, Asn to His), G to C at 639nt (213aa, Lys to Asn) of AB178712.1. Nucleotide sequence of AB178712.1 will be updated.
Vector backbone pcDNA3 (plasmid)
Size of vector backbone 5.4 kb
Selectable markers Amp^r (E. coli), Neo^r (mammalian cell)
Gene/insert name X. laevis CaM cDNA synthetic myosin light chain kinase peptide (M13) cDNA Aequorea victoria GFP cDNA
Reference of nucleotide sequence AB178712 (DDBJ accession number)
Depositor Miyawaki, Atsushi |
 
Other clones in our bank X. laevis (KEGG orthology K02183) | synthetic (KEGG orthology K00907) |

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.
Nagai T, Yamada S, Tominaga T, Ichikawa M, Miyawaki A. (2004) Expanded dynamic range of fluorescent indicators for Ca2+ by circularly permuted yellow fluorescent proteins. Proc. Natl. Acad. Sci. U S A. 101:10554-10559.
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.
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Remarks
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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB15135 YC3.60/pcDNA3 DNA solution

Ordering Information [in Japanese] [in English]

References and tips

Electronic file

Featured content

Featured content Dr. Miyawaki's resources (English text)
Featured content Calcium Ion Sensor Clones (English text)

References

original Miyawaki, A., Imaging intracellular free Ca2+ concentration using yellow cameleons. Cold Spring Harb. Protoc. 2013 (11). pii: pdb.prot078642 (2013). PMID 24184769.
original Nagai, T., Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins. Proc. Natl. Acad. Sci. U.S.A. 101 (29): 10554-10559 (2004). PMID 15247428.

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 RDB15135_A7H9p1-2.pdf

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

Primer: CMV-Forward RDB15135_A7H9a.seq
>D04498C2_A7H9_1_CMV-Forward_A05_02_ABI24.ab1
    1 ATCGGATCAA CTCCGCCCCA TTGACGCAAA TGGGCGGTAG GCGTGTACGG TGGGAGGTCT
   61 ATATAAGCAG AGCTCTCTGG CTAACTAGAG AACCCACTGC TTACTGGCTT ATCGAAATTA
  121 ATACGACTCA CTATAGGGAG ACCCAAGCTT GCGGCCGCCA CCATGGTGAG CAAGGGCGAG
  181 GAGCTGTTCA CCGGGGTGGT GCCCATCCTG GTCGAGCTGG ACGGCGACGT AAACGGCCAC
  241 AGGTTCAGCG TGTCCGGCGA GGGCGAGGGC GATGCCACCT ACGGCAAGCT GACCCTGAAG
  301 TTCATCTGCA CCACCGGCAA GCTGCCCGTG CCCTGGCCCA CCCTCGTGAC CACCCTGACC
  361 TGGGGCGTGC AGTGCTTCAG CCGCTACCCC GACCACATGA AGCAGCACGA CTTCTTCAAG
  421 TCCGCCATGC CCGAAGGCTA CGTCCAGGAG CGTACCATCT TCTTCAAGGA CGACGGCAAC
  481 TACAAGACCC GCGCCGAGGT GAAGTTCGAG GGCGACACCC TGGTGAACCG CATCGAGCTG
  541 AAGGGCATCG ACTTCAAGGA GGACGGCAAC ATCCTGGGGC ACAAGCTGGA GTACAACTAC
  601 ATCAGCCACA ACGTCTATAT CACCGCCGAC AAGCAGAAGA ACGGCATCAA GGCCCACTTC
  661 AAGATCCGCC ACAACATCGA GGACGGCAGC GTGCAGCTCG CCGACCACTA CCAGCAGAAC
  721 ACCCCCATCG GCGACGGCCC CGTGCTGCTG CCCGACAACC ACTACCTGAG CACCCAGTCC
  781 GCCCTGAGCA AAGACCCCAA CGAGAAGCGC GATCACATGG TCCTGCTGGA GTTCGTGACC
  841 GCCGCCCGCA TGCATGACCA ACTGACAGAA GAGCAGATTG CAGAGTTCAA AGAAGCCTTC
  901 TCATTATTCG ACAAGGGATG GGGACGGCAC CATCACCACA AAGGAACTTG GGCACCGTTA
  961 TGAGGTCGCT TGGGACAAAA CCCAACGGGA AGCAGAAATT GCAGGATATG ATCAAATGAA
 1021 GTCGATGCTG ATGGCAATGG GAACGATTTA CTTTCCTGAA TTTCTTACTA TGATGGGCTA
 1081 GAAAAAATGA AGGACACAGG ACAGCGAGAG AATCGAGAAG CATTCCGTGT TTTTGGAACA
1141 AGGAATTGGG AACCGCGGTC CT
//
Primer: BGH_rev2 RDB15135_A7H9b.seq
>D04498C2_A7H9_1_BGH_rev2_B05_05_ABI24.ab1
    1 CAATTCGGGG TTCAGGGAGG CACGGGGGAG GGGCAAACAA CAGATGGCTG GCAACTAGAA
   61 GGCACAGTCG AGGCTGATCA GCGAGCTCTA GCATTTAGGT GACACTATAG AATAGGGCCC
  121 TCTAGATGCA TGCTCGAGCG GCCGCCAGTG TGATGGATAT CTGCAGAATT CTTACTCGAT
  181 GTTGTGGCGG ATCTTGAAGT TGGCCTTGAT GCCGTTCTTC TGCTTGTCGG CGGTGATATA
  241 GACGTTGTGG CTGTTGTAGT TGTACTCCAG CTTGTGCCCC AGGATGTTGC CGTCCTCCTT
  301 GAAGTCGATG CCCTTCAGCT CGATGCGGTT CACCAGGGTG TCGCCCTCGA ACTTCACCTC
  361 GGCGCGGGTC TTGTAGTTGC CGTCGTCCTT GAAGAAGATG GTGCGCTCCT GGACGTAGCC
  421 TTCGGGCATG GCGGACTTGA AGAAGTCGTG CTGCTTCATG TGGTCGGGGT AGCGGGCGAA
  481 GCACTGCAGG CCGTAGCCCA GGGTGGTCAC GAGGGTGGGC CAGGGCACGG GCAGCTTGCC
  541 GGTGGTGCAG ATCAGCTTCA GGGTCAGCTT GCCGTAGGTG GCATCGCCCT CGCCCTCGCC
  601 GGACACGCTG AACTTGTGGC CGTTTACGTC GCCGTCCAGC TCGACCAGGA TGGGCACCAC
  661 CCCGGTGAAC AGCTCCTCGC CCTTGCTCAC CATGCCACCG CTGCCACCCT TGTACAGCTC
  721 GTCCATGCCG AGAGTGATCC CGGCGGCGGT CACGAACTCC AGCAGGACCA TGTGATCGCG
  781 CTTCTCGTTG GGGTCTTTGC TCAGGGCGGA CTGGTAGCTC AGGTAGTGGT TGTCGGGCAG
  841 CAGCACGGGG CCGTCGCCGA TGGGGGTGTT CTGCTGGTAG TGGTCGGCGA GCTGCACGCC
  901 GCCGTCCATG AGCTCCAGTG CCCCGGAGCT GGAGATCTTC TTGAACCGGT TGGCAGCGCT
  961 GACGGCAATG AAGTTTTTTC TTCCAGCGCC TCTTTCCCCC CCTTTTGCTG TCATCATTTG
 1021 TACAAACTCT TTCATAGTTT ACTTGGGCCC ATCACCCATC GATATCTGCT TTCCCTTATC
 1081 ATTTCATCAA CTTTCTTCAA TCTGTTAACT CTCCCGAGTT TGGTCATGAA CGTGACGTTA
1141 ACTGGAAAGC AGCGGCCTTG ATGTGTTATA GC
//
Primer: SV40pro_F RDB15135_A7H9c.seq
>15135_15135_A7H9_SV40pro_F_B02_02_ABI08.ab1
    1 CCCCCCTCCC TTTTTACCTT TTCAGAAAGT AGTGGAGGAG GCTTTTTTTG GAGGCCTAGG
   61 CTTTTGCAAA AAGCTCCCGG GAGCTTGTAT ATCCATTTTC GGATCTGATC AAGAGACAGG
  121 ATGAGGATCG TTTCGCATGA TTGAACAAGA TGGATTGCAC GCAGGTTCTC CGGCCGCTTG
  181 GGTGGAGAGG CTATTCGGCT ATGACTGGGC ACAACAGACA ATCGGCTGCT CTGATGCCGC
  241 CGTGTTCCGG CTGTCAGCGC AGGGGCGCCC GGTTCTTTTT GTCAAGACCG ACCTGTCCGG
  301 TGCCCTGAAT GAACTGCAGG ACGAGGCAGC GCGGCTATCG TGGCTGGCCA CGACGGGCGT
  361 TCCTTGCGCA GCTGTGCTCG ACGTTGTCAC TGAAGCGGGA AGGGACTGGC TGCTATTGGG
  421 CGAAGTGCCG GGGCAGGATC TCCTGTCATC TCACCTTGCT CCTGCCGAGA AAGTATCCAT
  481 CATGGCTGAT GCAATGCGGC GGCTGCATAC GCTTGATCCG GCTACCTGCC CATTCGACCA
  541 CCAAGCGAAA CATCGCATCG AGCGAGCACG TACTCGGATG GAAGCCGGTC TTGTCGATCA
  601 GGATGATCTG GACGAAGAGC ATCAGGGGCT CGCGCCAGCC GAACTGTTCG CCAGGCTCAA
  661 GGCGCGCATG CCCGACGGCG AGGATCTCGT CGTGACCCAT GGCGATGCCT GCTTGCCGAA
  721 TATCATGGTG GAAAATGGCC GCTTTTCTGG ATTCATCGAC TGTGGCCGGC TGGGTGTGGC
  781 GGACCGCTAT CAGGACATAG CGTTGGCTAC CCGTGATATT GCTGAAGAGC TTGGCGGCGA
  841 ATGGGCTGAC CGCTTCCTCG TGCTTTACGG GTATCGCCGC TCCCGATTCG CAGCGCATCG
  901 CCTTCTATCG CCTTCTTGAC GAGTTTCTTC TGAGCGGGAC TCTGGGGGTT TCGAAATGAC
  961 CGACCAAGCG ACGCCCAACC CTGCCCATCA CGAAGATTTC GATTCCACCC GCCCGCCCTT
 1021 TCTATGAAAA GGTTGGGGCT TTCGGAAATC GTTTTTTCCC GGGGACGCCC GGCCTGGGAT
 1081 GGATTCCTTC CAGCCGCGGG GAATCTCCAT GCCTTGGGAA GTTCTTTCGC CCAACCCCCC
1141 AATCTTGTTT AAATTGCCAG CTTTAATGAT GGGTTTAACA AAATTAAAAA GGCC
//

Please visit Sequencing and PCR primers for primer information.


References

References

original Miyawaki, A., Imaging intracellular free Ca2+ concentration using yellow cameleons. Cold Spring Harb. Protoc. 2013 (11). pii: pdb.prot078642 (2013). PMID 24184769.
original Nagai, T., Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins. Proc. Natl. Acad. Sci. U.S.A. 101 (29): 10554-10559 (2004). PMID 15247428.

2019.03.19

GNP_filter3_RDBDEP_html_190316.pl