RDB15139 - Flash pericam/pcDNA3

Please review the QC test results indicated by check

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Flash pericam/pcDNA3 (#RDB15139)

Calcium ion sensor clone Flash pericam having a circularly permuted YFP and a CaM.

Clone info.	Calcium ion sensor clone Flash pericam having a circularly permuted YFP and a CaM. Better at 28-30oC than 37oC. Refer Fig.1 of Nagai, T. Proc. Natl. Acad. Sci. U.S.A. 98 (6): 3197-3202, 2001 for its composition.
Vector backbone	pcDNA3 (plasmid)
Selectable markers	Amp^r (E. coli), Neo^r (mammalian cells)
Gene/insert name	X. laevis CaM cDNA synthetic myosin light chain kinase peptide (M13) cDNA Aequorea victoria GFP cDNA
Depositor\|Developer	Miyawaki, Atsushi \|

Sequence (partial)	RDB15139pka02.seq provided by the depositor

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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Order form [Credit Card Payment] [Bank Transfer Payment] [Example of order form ]
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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. 98 (6): 3197-3202 (2001).

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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 ] 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. 98 (6): 3197-3202 (2001).

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。
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. 98 (6): 3197-3202 (2001).
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.)
RDB15139	Flash pericam/pcDNA3	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 Flash pericam/pcDNA3 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15139).

Reference section:

Nagai, T., Sawano, A., Park, E.S., Miyawaki, A., Circularly permuted green fluorescent proteins engineered to sense Ca2+. Proc. Natl. Acad. Sci. U.S.A. 98 (6): 3197-3202 (2001). PMID 11248055. [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	RDB15139_A7H9p1-1.pdf

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

Primer: CMV_Forward (Pr0016) Region: CMV pro, T7 pro, insert 5' Sequence file: RDB15139_A7H9a.seq
>D04498C6_A7H9_1_CMV-Forward_B02_05_ABI24.ab1 1 NNNNNNNNNN NNNNNNNTNG NNGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA 61 TAAGCAGAGC TCTCTGGCTA ACTAGAGAAC CCACTGCTTA CTGGCTTATC GAAATTAATA 121 CGACTCACTA TAGGGAGACC CAAGCTTGCG GCCGCCACCA TGGTGAAGAG GCGCTGGAAG 181 AAAAACTTCA TTGCCGTCAG CGCTGCCAAC CGGTTCAAGA AGATCTCCAG CTCCGGGGCA 241 CTGGGGTCTG CAGGCTACAA CAGCCACAAC GTCTATATCA TGGCCGACAA GCAGAAGAAC 301 GGCATCAAGG CCAACTTCAA GATCCGCCAC AACATCGAGG ACGGCGGCGT GCAGCTCGCC 361 GACCACTACC AGCAGAACAC CCCCATCGGC GACGGCCCCG TGCTGCTGCC CGACAACCAC 421 TACCTGAGCC ACCAGTCCGC CCTGAGCAAA GACCCCAACG AGAAGCGCGA TCACATGGTC 481 CTGCTGGAGT TCGTGACCGC CGCCGGGATC ACTCTCGGCA TGGACGAGCT GTACAAGGGT 541 GGCAGCGGTG GCATGGTGAG CAAGGGCGAG GAGCTGTTCA CCGGGGTGGT GCCCATCCTG 601 GTCGAGCTGG ACGGCGACGT AAACGGCCAC AAGTTCAGCG TGTCCGGCGA GGGCGAGGGC 661 GATGCCACCT ACGGCAAGCT GACCCTGAAG TTCATCTGCA CCACCGGCAA GCTGCCCGTG 721 CCCTGNCCCA CCCTCGTGAN CACCTTCGGC TACGGCCTGA AGTGCTTCGC CCGCTACCCC 781 GACCACATGA AGCAGCACGA CTTCTTCAAG TCCGCCATGC CCGAAGGCTA CGTCCAGGAN 841 CGCACCATCT TCTTCNAGGA CGACGGCAAC TACAAGANCC GCGCCGANGT GAANTNCGAG 901 GGCGACACCC NGGNGAACCG CATCGAGCTN AANGGCATCG NNTTCNAGGA NGNNNGNANC 961 NTCCNNGGGN NNNNNCNNGN NNTANNNCNG NNACNNGNNN GNNNNNNCNN NNNNNNNN //
Primer: BGH_rev2 (Pr0606) Region: insert 3' Sequence file: RDB15139_A7H9b.seq
>D04498C6_A7H9_1_BGH_rev2_D06_12_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNANNN NNNNNNGGCT GGCAANNNNN 61 NNGNACAGTC GAGGNNNNNN NNNNNNNCTC TAGCATTTAG GTGACACTAT AGAATAGGGC 121 CCNCTANATG CATGCTCGAG CGGCCGCCAG TGTGATGGAT ATCTGCAGAA TTCTTACTTT 181 GCTGTCATCA TTTGTACAAA CTCTTCATAG TTTACTTGGC CATCACCATC GATATCTGCT 241 TCCCTTATCA TTTCATCAAC TTCTTCATCT GTTAACTTCT CCCCGAGGTT TGTCATGACG 301 TGACGTAACT GAGCAGCGCT GATGTAGCCG TTCCCATCCT TGTCAAAAAC ACGGAATGCT 361 TCTCGGATTT CCTCTTCGCT GTCTGTGTCC TTCATTTTTC TAGCCATCAT AGTAAGAAAT 421 TCAGGAAAGT AAATCGTTCC ATTGCCATCA GCATCGACTT CATTGATCAT ATCCTGCAAT 481 TCTGCTTCCG TTGGGTTTTG TCCAAGCGAC CTCATAACGG TGCCAAGTTC CTTTGTGGTG 541 ATGGTGCCGT CCCCATCCTT GTCGAATAAT GAGAAGGCTT CTTTGAACTC TGCNNNNNNC 601 TCTTCTGTCA NTTGGTCATG CATCCCNGNN NNGTTGTACT CCAGCTTGTG CCCCANNNTG 661 TTGCCGTCCT CCTTGAANTC GATGCCCTTC AGCTCGATGC GGTTCNCCAG GGTGTCGCCC 721 TCNAACNNNN NNTCNGCGCG GGTCTTGTAN TTGNNNNCGT CCTTGAAGAA GANGGGTGCG 781 CTCCTGGACN NNNNNCTTCN GGNNNTGGCG GNACTTGAAG AANNTCGTGC TGCTTCATGT 841 GGTCNGGGGT ANCGGGCGAA GCACTTCANG NCNGNANCCN NNNNGNGGTC ACNNANGGTG 901 GGCNNNGGCA CGGGCAGCTT GCCGGTGGTG CANNNGAAAC TTCNNGGGTC NNCNNGNCNG 961 NANGNNNCNN CNCCCTNNNC CTCNCNNNAN NNNCTGAACT TGNGGNCNNN TTTANNNNNN 1021 NNNGNNNNNN NNNN //
Primer: EGFP-N_F (Pr0649) Region: insert mid Sequence file: RDB15139_A7H9c.seq
>D04498C6_A7H9_1_EGFP-N_F_G07_19_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN CNNNGGCGNN GCNNCCTACG GCAAGCTGNN CCTGAAGTTC 61 ATCTGCACCA CCGGCAAGCT GCCCGTGCCC TGGCCCACCC TCGTGACCAC CTTCGGCTAC 121 GGCCTGAAGT GCTTCGCCCG CTACCCCGAC CACATGAAGC AGCACGACTT CTTCAAGTCC 181 GCCATGCCCG AAGGCTACGT CCAGGAGCGC ACCATCTTCT TCAAGGACGA CGGCAACTAC 241 AAGACCCGCG CCGAGGTGAA GTTCGAGGGC GACACCCTGG TGAACCGCAT CGAGCTGAAG 301 GGCATCGACT TCAAGGAGGA CGGCAACATC CTGGGGCACA AGCTGGAGTA CAACGGTACC 361 GGGATGCATG ACCAACTGAC AGAAGAGCAG ATTGCAGAGT TCAAAGAAGC CTTCTCATTA 421 TTCGACAAGG ATGGGGACGG CACCATCACC ACAAAGGAAC TTGGCACCGT TATGAGGTCG 481 CTTGGACAAA ACCCAACGGA AGCAGAATTG CAGGATATGA TCAATGAAGT CGATGCTGAT 541 GGCAATGGAA CGATTTACTT TCCTGAATTT CTTACTATGA TGGCTAGAAA AATGAAGGAC 601 ACAGACAGCG AAGAGGAAAT CCGAGAAGCA TTCCGTGTTT TTGACAAGGA TGGGAACGGC 661 TACATCAGCG CTGCTCAGTT ACGTCACGTC ATGACAAACC TCGGGGAGAA GTTAACAGAT 721 GAAGAAGTTG ANGAAATGAT AANGGAAGCA GATATCGATG GTGATGGCCA AGTAAACTAT 781 GAAGAGTTTG TACNAATGNN GACAGCAAAG TAAGAATTCT GCAGATATCC ATCACACTGG 841 CGGNNGCTCN NNCATGNATC TANAGGGNCC TATTCTATAN TGTCANNNAA ATGCTANNNC 901 TCGNNNNNCA NCNNCNNNNG NNCNNTCTAN NNNNNANCNN TCTNNNNNTN GNCCNNNCCC 961 NNNNNNNNNC NNTNNNNCNN NNNNNNNN //
Primer: SV40pro_ori_F (Pr0731) Region: NeoR Sequence file: RDB15139_A7H9d.seq
>D04498C6_A7H9_1_SV40pro_ori_F_H07_22_ABI24.ab1 1 NNNNNNNNNN NNNGNCGCCT CNGCCTCTGN NCTATTCNNG AAGTAGTGAG GAGGCTTTTT 61 TGGAGGCCTA GGCTTTTGCA AAAAGCTCCC GGGAGCTTGT ATATCCATTT TCGGATCTGA 121 TCAAGAGACA GGNTGAGGAT CGTTTCGCAT GATTGAACAA GATGGATTGC ACGCAGGTTC 181 TCCGGCCGCT TGGGTGGAGA GGCTATTCGG CTATGACTGG GCACAACAGA CAATCGGCTG 241 CTCTGATGCC GCCGTGTTCC GGCTGTCAGC GCAGGGGCGC CCGGTTCTTT TTGTCAAGAC 301 CGACCTGTCC GGTGCCCTGA ATGAACTGCA GGACGAGGCA GCGCGGCTAT CGTGGCTGGC 361 CACGACGGGC GTTCCTTGCG CAGCTGTGCT CGACGTTGTC ACTGAAGCGG GAAGGGACTG 421 GCTGCTATTG GGCGAAGTGC CGGGGCAGGA TCTCCTGTCA TCTCACCTTG CTCCTGCCGA 481 GAAAGTATCC ATCATGGCTG ATGCAATGCG GCGGCTGCAT ACGCTTGATC CGGCTACCTG 541 CCCATTCGAC CACCAAGCGA AACATCGCAT CGAGCGAGCA CGTACTCGGA TGGAAGCCGG 601 TCTTGTCGAT CAGGATGATC TGGACGAAGA GCATCAGGGG CTCGCGCCAG CCGAACTGTT 661 CGCCAGGCTC AAGGCGCGCA TGCCCGACGG CGANGNNCTC GTCGTGACCC ATGGCGATGC 721 CTGCTTGCCG AATATCATGG TGGANNATGG CCGCTTTTCT GGANTCATCG ACTGTGGCCG 781 GCTGNNTGTG GCGGANCGCT ATCANGNCAT ANCGTTGGCT ANCCGTGATA TTGCTGAAGA 841 NCTTGGCGGC GAATGGGCTG ANCGCTTCCT CGNNCTTTAC NGNANCGCNN CNCCNNATTN 901 NCANCNNATC NNCNTTCNNN CGNCNTTNNN GNCNANNTNN NNNNNNNN //

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original	Nagai, T., Circularly permuted green fluorescent proteins engineered to sense Ca2+. Proc. Natl. Acad. Sci. U.S.A. 98 (6): 3197-3202 (2001). PMID 11248055. [link to RRC of NBRP]