RDB15137 - YC3.60pm/pRSETB

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YC3.60pm/pRSETB (#RDB15137)

Yellow cameleon calcium ion sensor clone YC3.60pm wiht plasma membrane localization tag.

Clone info.	Calcium ion sensor clone YC3.60 Yellow cameleon having plasma membrane localization tag, E104Q substitution in the 3rd Ca2+ binding site of CaM and the circular permutated cp173Venus. Bacterial expression plasmid.
Comment	The CAAX box of Ki-Ras was fused to the C terminus of YC3.60 through a linker sequence (GTGGSGGGTGGSGGGT).
Vector backbone	pRSETB (plasmid)
Size of vector backbone	2.9 kb
Selectable markers	Amp^r (E. coli)
Growth conditions	LB + Amp, 37oC
Gene/insert name	X. laevis CaM cDNA synthetic myosin light chain kinase peptide (M13) cDNA Aequorea victoria GFP cDNA
Reference of insert sequence	AB178714 (DDBJ accession number)
Depositor\|Developer	Miyawaki, Atsushi \|

Sequence (full)	RDB15137zzt01.seq compiled by us

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.

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Terms and conditions for distribution When publishing the results obtained using the BIOLOGICAL RESOURCE, a citation of literature specified by Dr. Atsushi Miyawaki or an acknowledgement to Dr. Miyawaki is requested.
Nagai, T. et al., Proc. Natl. Acad. Sci. U S A. 101:10554-10559 (2004).

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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 ] MTA, for use for for-profit purpose [Word] Please visit Information of Request for Distribution.[link]
Terms and conditions for distribution	When publishing the results obtained using the BIOLOGICAL RESOURCE, a citation of literature specified by Dr. Atsushi Miyawaki or an acknowledgement to Dr. Miyawaki is requested. Nagai, T. et al., Proc. Natl. Acad. Sci. U S A. 101:10554-10559 (2004).

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 ] 提供同意書 (MTA, 営利目的用)[Word] 手続きの概要は、「提供申込みについて[link]」をご覧ください。
MTAに書く使用条件	When publishing the results obtained using the BIOLOGICAL RESOURCE, a citation of literature specified by Dr. Atsushi Miyawaki or an acknowledgement to Dr. Miyawaki is requested. Nagai, T. et al., Proc. Natl. Acad. Sci. U S A. 101:10554-10559 (2004).
備考	提供条件は英文のまま記載してください。

Catalog #	Resource name	Shipping form	Fee (non-profit org.)
RDB15137	YC3.60pm/pRSETB	DNA solution	JPY 9,460 (not-for-profit academic purpose) JPY 18,920 (for-profit-research 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 YC3.60pm/pRSETB was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15137).

Reference section:

Miyawaki, A., Nagai, T., Mizuno, H., Imaging intracellular free Ca2+ concentration using yellow cameleons. Cold Spring Harb. Protoc. 2013 (11). pii: pdb.prot078642 (2013). PMID 24184769. [link to RRC of NBRP]

Nagai, T., Yamada, S., Tominaga, T., Ichikawa, M., Miyawaki, A., 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. [link to RRC of NBRP]

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	RDB15137_A7H9p1-1.pdf

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

Primer: lacIpro-B_F (Pr0724) Region: T7 pro, RBS, 6xHis, T7 tag, Xpress tag, insert 5' Sequence file: RDB15137_A7H9a.seq
>15137_15137_B0KrF_lacIpro-B_F_A07_01_ABI24.ab1 1 NNNNNNNNNN NNNNNNCGCA NCGNNNCNGT GNGCGANGAA GCGGAAGAGC GCCCAATACG 61 CAAACCGCCT CTCCCCGCGC GTTGGCCGAT TCATTAATGC AGGATCTCGA TCCCGCGAAA 121 TTAATACGAC TCACTATAGG GAGACCACAA CGGTTTCCCT CTAGAAATAA TTTTGTTTAA 181 CTTTAAGAAG GAGATATACA TATGCGGGGT TCTCATCATC ATCATCATCA TGGTATGGCT 241 AGCATGACTG GTGGACAGCA AATGGGTCGG GATCTGTACG ACGATGACGA TAAGGATCCC 301 ATGGTGAGCA AGGGCGAGGA GCTGTTCACC GGGGTGGTGC CCATCCTGGT CGAGCTGGAC 361 GGCGACGTAA ACGGCCACAG GTTCAGCGTG TCCGGCGAGG GCGAGGGCGA TGCCACCTAC 421 GGCAAGCTGA CCCTGAAGTT CATCTGCACC ACCGGCAAGC TGCCCGTGCC CTGGCCCACC 481 CTCGTGACCA CCCTGACCTG GGGCGTGCAG TGCTTCAGCC GCTACCCCGA CCACATGAAG 541 CAGCACGACT TCTTCAAGTC CGCCATGCCC GAAGGCTACG TCCAGGAGCG TACCATCTTC 601 TTCAAGGACG ACGGCAACTA CAAGACCCGC GCCGAGGTGA AGTTCGAGGG CGACACCCTG 661 GTGAACCGCA TCGAGCTGAA NGGCATCGAC TTCAAGGANG ACGGCAACAT CCTGGGGCAC 721 AAGCTGGAGT ACAACTACAT CAGCCACAAC GTCTATATCA CCGCCGANAA GCAGAAGAAC 781 GGCATCAAGG CCCACTTCAA GATCCGCCAC AACATCGAGG ANGGCAGCGN GCAGCTCGCC 841 NACCACTACC AGCAGAACAC CCNCATCGGC GANGGCCCCG NNCTGCTGCC CGACAACCAC 901 TACCNGANCA CCCNNNNCGC CCNNNNCAAA GANCNNNANG ANAANCGCGN NNNCNNNGNC 961 CNGNNNGNNN NNNNNNN //
Primer: T7ter_R1 (Pr0798) Region: insert 3' Sequence file: RDB15137_A7H9b.seq
>15137_15137_B0KrF_T7ter_R1_B07_04_ABI24.ab1 1 NNNNNNNNNN NNNNNNNGNN NNTNNNTGCT NNGCGGTGGC AGCAGCCNAC TCAGCTTCCT 61 TTCGGGCTTT GTTAGCAGCC GGATCAAGCT TCGAATTCTT ACATAATTAC ACACTTTGTC 121 TTTGACTTCT TTTTCTTCTT TTTACCATCT TTGCTCATCT TGGTACCGGA GCCGCCGCTG 181 CCGCCGGTAC CGGAGCCGCC GCTGCCGCCG GTACCCTCGA TGTTGTGGCG GATCTTGAAG 241 TTGGCCTTGA TGCCGTTCTT CTGCTTGTCG GCGGTGATAT AGACGTTGTG GCTGTTGTAG 301 TTGTACTCCA GCTTGTGCCC CAGGATGTTG CCGTCCTCCT TGAAGTCGAT GCCCTTCAGC 361 TCGATGCGGT TCACCAGGGT GTCGCCCTCG AACTTCACCT CGGCGCGGGT CTTGTAGTTG 421 CCGTCGTCCT TGAAGAAGAT GGTGCGCTCC TGGACGTAGC CTTCGGGCAT GGCGGACTTG 481 AAGAAGTCGT GCTGCTTCAT GTGGTCGGGG TAGCGGGCGA AGCACTGCAG GCCGTAGCCC 541 AGGGTGGTCA CGAGGGTGGG CCAGGGCACG GGCAGCTTGC CGGTGGTGCA GATCAGCTTC 601 AGGGTCAGCT TGCCGTANGT GGCATCGCCC TCGCCCTCGC CGGACACGCT GAACTTGTGG 661 CCGTTTACGT CGCCGTCCAG CTCGACCAGG ATGGGCACCA CCCCGGTGAA CAGCTCCTCG 721 CCCTTGCTCA CCATGCCACC GCTGCCACCC TTGTACAGCT CGTCCATGCC GANAGTGATC 781 CCGGCGGCGG TCACGAACTC CAGCANGNNC ATGTGATCGC GCTTCTCGTT GGGGTCTTTG 841 CTCANGGCGG NNNGGTANCT CANGTAGTGN TTGTCNGGCA GCANCACGGG GCCNNCGCCN 901 ATGGGGNNGT TCTGCNNGNA NTGGTCNGNN ANCNGCACGN CNNNNNNCAT NANCNNCNNN 961 NNCCCNNNNN NNGNNNNNNN NNN //
Primer: FP-N_F1 (Pr0920) Region: insert mid Sequence file: RDB15137_A7H9c.seq
>15137_15137_B0KrF_FP-N_F1_C07_07_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNCN TGNAGCAGCA CGACTTCTTC NAGTCCGCCA 61 TGCCCGAAGG CTACGTCCAG GAGCGTACCA TCTTCTTCAA GGACGACGGC AACTACAAGA 121 CCCGCGCCGA GGTGAAGTTC GAGGGCGACA CCCTGGTGAA CCGCATCGAG CTGAAGGGCA 181 TCGACTTCAA GGAGGACGGC AACATCCTGG GGCACAAGCT GGAGTACAAC TACATCAGCC 241 ACAACGTCTA TATCACCGCC GACAAGCAGA AGAACGGCAT CAAGGCCCAC TTCAAGATCC 301 GCCACAACAT CGAGGACGGC AGCGTGCAGC TCGCCGACCA CTACCAGCAG AACACCCCCA 361 TCGGCGACGG CCCCGTGCTG CTGCCCGACA ACCACTACCT GAGCACCCAG TCCGCCCTGA 421 GCAAAGACCC CAACGAGAAG CGCGATCACA TGGTCCTGCT GGAGTTCGTG ACCGCCGCCC 481 GCATGCATGA CCAACTGACA GAAGAGCAGA TTGCAGAGTT CAAAGAAGCC TTCTCATTAT 541 TCGACAAGGA TGGGGACGGC ACCATCACCA CAAAGGAACT TGGCACCGTT ATGAGGTCGC 601 TTGGACAAAA CCCAACGGAA GCAGAATTGC AGGATATGAT CAATGAAGTC GATGCTGATG 661 GCAATGGAAC GATTTACTTT CCTGAATTTC TTACTATGAT GGCTAGAAAA ATGAAGGACA 721 CAGACAGCGA AGAGGAAATC CGAGAAGCAT TCCGTGTTTT TGACAAGGAT GGGAACGGCT 781 ACATCAGCGC TGCTCAGTTA CGTCACGTCA TGACAAACCT CGGGGANAAG TTAACAGATG 841 AAGAAGTTGA TGAAATGATA AGGNAAGCAG ATATCGATGG TGATGGNCNA GTAAACTATG 901 AAGAGTTTGT ACAAATGATG ACAGCAAANG GGGGGAAGAN GNGCNNGNNN AAAAAACTNC 961 NNNNNNNNCA GCGCNNNNNN CCGGNNCNNN NANNNNNNNC NNNNNNNGGN NNNNNNNNN //
Primer: YC4.6-M_F (Pr0721) Region: insert mid Sequence file: RDB15137_A7H9d.seq
>15137_15137_B0KrF_YC4.6-M_F_D07_10_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNC NTNNNGANGT CGCTTGGNNN AAACCCAACG GAAGCAGAAT 61 TGCAGGATAT GATCAATGAA GTCGATGCTG ATGGCAATGG AACGATTTAC TTTCCTGAAT 121 TTCTTACTAT GATGGCTAGA AAAATGAAGG ACACAGACAG CGAAGAGGAA ATCCGAGAAG 181 CATTCCGTGT TTTTGACAAG GATGGGAACG GCTACATCAG CGCTGCTCAG TTACGTCACG 241 TCATGACAAA CCTCGGGGAG AAGTTAACAG ATGAAGAAGT TGATGAAATG ATAAGGGAAG 301 CAGATATCGA TGGTGATGGC CAAGTAAACT ATGAAGAGTT TGTACAAATG ATGACAGCAA 361 AGGGGGGGAA GAGGCGCTGG AAGAAAAACT TCATTGCCGT CAGCGCTGCC AACCGGTTCA 421 AGAAGATCTC CAGCTCCGGG GCACTGGAGC TCATGGACGG CGGCGTGCAG CTCGCCGACC 481 ACTACCAGCA GAACACCCCC ATCGGCGACG GCCCCGTGCT GCTGCCCGAC AACCACTACC 541 TGAGCTACCA GTCCGCCCTG AGCAAAGACC CCAACGAGAA GCGCGATCAC ATGGTCCTGC 601 TGGAGTTCGT GACCGCCGCC GGGATCACTC TCGGCATGGA CGAGCTGTAC AAGGGTGGCA 661 GCGGTGGCAT GGTGAGCAAG GGCGAGGANC TGTTCACCGG GGTGGTGCCC ATCCTGGTCG 721 ANCTGGACGG CGACGTAAAC GGCCACAAGT TCAGCGTGTC CGGCGANGGC GANGGCGATG 781 CCACCTACNG CAAGCTGACC CTGAAGCTGA TCTGCACCAC CGGCAAGCTG CCCGTGCCCT 841 NNCCCACCCT CNNGANCACC CTGGGCTACN GNCNGCANNG CTTCGCCCGC TANCCCGNNC 901 ACATGANNCA NCNCGANTTC NNCNAGNCCG NCNTGNCCGN ANNNNACNNN CAGGNNCGCA 961 NNATCNTNNN NNNANNNNNN NNNGNNNNNN NNNN //
Primer: EYFP-C_F (Pr0412) Region: insert mid Sequence file: RDB15137_A7H9e.seq
>15137_15137_B0KrF_EYFP-C_F_E07_13_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNCGCGNNC NCNTGGTCCT GCTGGNNTTC GTGNCCGCCG 61 CCGGGNTCAC TCTCGGCATG GACGAGCTGT ACAAGGGTGG CAGCGGTGGC ATGGTGAGCA 121 AGGGCGAGGA GCTGTTCACC GGGGTGGTGC CCATCCTGGT CGAGCTGGAC GGCGACGTAA 181 ACGGCCACAA GTTCAGCGTG TCCGGCGAGG GCGAGGGCGA TGCCACCTAC GGCAAGCTGA 241 CCCTGAAGCT GATCTGCACC ACCGGCAAGC TGCCCGTGCC CTGGCCCACC CTCGTGACCA 301 CCCTGGGCTA CGGCCTGCAG TGCTTCGCCC GCTACCCCGA CCACATGAAG CAGCACGACT 361 TCTTCAAGTC CGCCATGCCC GAAGGCTACG TCCAGGAGCG CACCATCTTC TTCAAGGACG 421 ACGGCAACTA CAAGACCCGC GCCGAGGTGA AGTTCGAGGG CGACACCCTG GTGAACCGCA 481 TCGAGCTGAA GGGCATCGAC TTCAAGGAGG ACGGCAACAT CCTGGGGCAC AAGCTGGAGT 541 ACAACTACAA CAGCCACAAC GTCTATATCA CCGCCGACAA GCAGAAGAAC GGCATCAAGG 601 CCAACTTCAA GATCCGCCAC AACATCGAGG GTACCGGCGG CAGCGGCGGC TCCGGTACCG 661 GCGGCAGCGG CGGCTCCGGT ACCAAGATGA GCAAAGATGG TAAAAAGAAG AAAAAGAAGT 721 CAAAGACAAA GTGTGTAATT ATGTAAGAAN TCGAANCTTG ATCCGGCTGC TAACAAAGCC 781 CGAAAGGAAG CTGANTTGGC TGCTGCCACC GCTNANCAAT AACTAGCATA ACCCCTNGGG 841 GCCTCTAAAC GGGTCTTGAN GGNTTTTTTT GCTGAAAGGA NGAACTATAT CCGGATCTGG 901 CGNAATANCG AANANGCCCG CANCNATCGC CCNTCCNNAN AGTNGCNCAN CCNNNNNGGN 961 NAANNGNANG CGNCNNGNAN CGGNNNNNNN NNCNNNGNNN NNNNNGNNNN NNNNNN //

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original	Miyawaki, A., Imaging intracellular free Ca2+ concentration using yellow cameleons. Cold Spring Harb. Protoc. 2013 (11). pii: pdb.prot078642 (2013). PMID 24184769. [link to RRC of NBRP]
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. [link to RRC of NBRP]