RDB16750 - pCAGGS-ChRGR(ER)-Venus

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pCAGGS-ChRGR(ER)-Venus (#RDB16750)

Expression vector of organelle optogenetics channelrhodopsin.

Clone info.	Mammarian expression vector. Channelrhodopsin-green receiver with an ER-retention motif (ChRGR(ER)), tagged with Venus at C-terminus, CAG promoter. Codon optimized for human. To generate a ChRGR(ER), a cDNA encoding Gln4765-Ile4866 of mouse ryanodine receptor 2 (NM_023868) was inserted in-frame between ChRGR and Venus using the AgeI restriction site.
Comment	Expression was confirmed by the depositor with immunostaining.
Vector backbone	pCAGGS (Plasmid)
Size of vector backbone	4.8 kb
Selectable markers	Amp^r (E. coli).
Growth conditions	LB + Amp, 37oC.
Gene/insert name	Chlamydomonas reinhardtii acop1 (channelopsin-1) cDNA Chlamydomonas reinhardtii cop4 (acop2) cDNA
Reference of insert sequence	LC415030 (DDBJ accession number)
Depositor\|Developer	Ishizuka, Toru \|

Distribution information

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Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested (Asano, T., Front. Neurosci. 12: 561, 2018). For use of the BIOLOGICAL RESOURCE by a for-profit organization, the RECIPIENT must reach agreement on terms and conditions of use of it with DEPOSITOR and must obtain a prior written consent from the DEPOSITOR.
Additional terms and conditions:
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., Nat. Biotechnol. 20: 87-90 (2002).

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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	In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested (Asano, T., Front. Neurosci. 12: 561, 2018). For use of the BIOLOGICAL RESOURCE by a for-profit organization, the RECIPIENT must reach agreement on terms and conditions of use of it with DEPOSITOR and must obtain a prior written consent from the DEPOSITOR. Additional terms and conditions: 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., Nat. Biotechnol. 20: 87-90 (2002).

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	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 (Asano, T., Front. Neurosci. 12: 561, 2018)。営利機関の利用希望者は、事前に利用条件等につき寄託者と合意し、提供承諾を得ること。付加的使用条件： 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., Nat. Biotechnol. 20: 87-90 (2002).

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

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 (Asano, T., Front. Neurosci. 12: 561, 2018)。営利機関の利用希望者は、事前に利用条件等につき寄託者と合意し、提供承諾を得ること。
付加的使用条件：
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., Nat. Biotechnol. 20: 87-90 (2002).

Catalog #	Resource name	Shipping form	Fee (non-profit org.)
RDB16750	pCAGGS-ChRGR(ER)-Venus	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 pCAGGS-ChRGR(ER)-Venus was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB16750).

Reference section:

Asano, T., Igarashi, H., Ishizuka, T., Yawo, H., Organelle Optogenetics: Direct Manipulation of Intracellular Ca2+ Dynamics by Light. Front. Neurosci. 12: 561 (2018). PMID 30174581. [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	RDB16750_A8Jnp1-3.pdf

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

Primer: pAxCA_F1 (Pr0164) Region: kozak seq,insert 5'(ChRGR(ER) 5') Sequence file: RDB16750_A8Jnf.seq
>D05919A2_A8Jn_2_pAxCA-F1_G03_21_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNT GCCTTCTTCT TTTTCCTACN GCTCCTGGGC AACGTGCTGG 61 TTATTGTGCT GTCTCATCAT TTTGGCAAAG AATTCGCCAC CATGGCTCGG AGACCTTGGC 121 TGCTGGCACT GGCTCTGGCT GTCGCTCTGG CCGCTGGCTC TGCTGGGGCT TCTACTGGAT 181 CTGACGCAAC AGTGCCCGTC GCTACTCAGG ACGGACCTGA TTACGTGTTC CACAGGGCAC 241 ATGAGCGCAT GCTGTTTCAG ACATCTTATA CTCTGGAAAA CAATGGCAGT GTCATCTGCA 301 TTCCCAACAA TGGGCAGTGC TTCTGTCTGG CATGGCTGAA GTCTAACGGC ACCAATGCCG 361 AGAAACTGGC CGCTAACATC CTGCAGTGGA TTACATTCGC CCTGTCCGCT CTGTGCCTGA 421 TGTTTTACGG ATATCAGACC TGGAAGTCTA CATGTGGCTG GGAGGAAATC TACGTGGCTA 481 CCATCGAGAT GATCAAGTTC ATCATTGAAT ACTTCCACGA GTTTGACGAA CCTGCCGTGA 541 TCTATAGCTC CAACGGGAAT AAGACTGTCT GGCTGAGATA CGCTGAGTGG CTGCTGACCT 601 GTCCAGTGAT CCTGATTCAT CTGAGCAACC TGACTGGCCT GGCCAACGAC TATAATAAGA 661 GAACCATGGG GCTGCTGGTG AGTGATATCG GAACAATTGT CTGGGGCACC ACAGCAGCCC 721 TGTCAAAAGG CTATGTGCGC GTCATCTTCT TTCTGATGGG CCTGTGCTAC GGGATCTACA 781 CATTCTTTAA TGCTGCAAAG GTGTACATCG NGGCATATCA CACAGTCCNT AAAGGGCGGT 841 GTAGACAGGT GGTCACTGGA ATGGCCTGNC TGTNCTTTGT GTNCTGGGGG ANGTTNCNNA 901 TCCNGNTTCT GCTGGGNNCN GAANNNNNCN GCCNNNNNNA NNNGTTTNNN NCAGCCNTCG 961 CNNATNCNNN TCTGNNNNNN NNNN //
Primer: Venus-N_R (Pr0341) Region: insert mid(Venus 5',ChRGR(ER) 3') Sequence file: RDB16750_A8Jng.seq
>D05919A2_A8Jn_2_venus-N_R_H03_24_ABI24.ab1 1 NNNNNNNNNN NNCNNNNTTG TGGNCGTTNN NGTCGCCGTC CNGCTCGACC AGGNTGGGCA 61 CCACCCCGGT GAACAGCTCC TCGCCCTTGC TCACCATGGT GGCGACCGGT ATGATCAAAC 121 CTTGTATGAT AGCCAAAAGG ATGACAATCA CGAAGAAGAA GAATGTGATG TCAAAGATGA 181 TGCGGTAGAT CTCATACTCG TCTCCTGCTG GGTCTTCAAT TTCATCACCA ATGCCCCCTC 241 CAGCACGGAC GCCCACATAC ATGTGGAACA TGTAACACGT CAGCATGTCA TCACACTTCA 301 TGTCTGGCGT GTCACCATCC TCACTTTTAT TGTAGAATTT TCGGAAGAAA TTGAACGCCA 361 CGACAGTGTA CAGGTACACA ACAACAGCCA ATAAGCCAAC AGTTAACACG AGCTGTACCG 421 GTGGATCCTT CCCAGTCCCT TTGTTCACGG CTCCTGCCTC GGCTTCATCT TCCACCAGTG 481 TTTCCACCTC AATTTCGGTC CCGCCGATAT TCAGTTTGGT AGTCTTCCGA ATATCGCCAT 541 GAATCAGGAT ATGCTCGTGG ATCAGCACTC GCAGGTAGTG TCCCAGCAGC CCCCAGCAGT 601 TTTTGCTGGC CAGGTCCAGA ATTGCATGAG CGATGGCTGA ATTAAACTGG TTAATGTGGC 661 CGAATCCTTC GGGTCCCAGC AGAAACAGGA TTGGGAACAT CCCCCAGGAC ACAAAGAACA 721 GCCAGGCCAT TCCAGTGACC ACCTGTCTAC ACCGCCCTTT ANGGACTGTG TGATATGCCT 781 CGATGTACAC CTTTGCAGCA TTNAAGAATG TGTAGATCCC GTANCACANG CCCATCAGAA 841 AGAANANGAC GCGCACATAG CCTTTTGACA GGGCTGCTGN GGTGCCCCAG ANAATTGTNN 901 CGANATCACT CACCANCAGC CCCANNNTTC TNTNATTATA NNCGTNNNNC ANGCNANNNN 961 NNTNGNNNAN NNGNNNNNNN ATCANNNGNN NNNGNNNNCN NNNNNNNNN //
Primer: reverse (Pr0099) Region: beta globin pA,insert 3'(Venus 3') Sequence file: RDB16750_A8Jnh.seq
>D05919A2_A8Jn_2_reverse_H02_23_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNNNNNCNT AAGAGAAGAG GGNNAGCTAT GACTGGGAGT 61 AGTCAGGAGA GGAGGAAAAA TCTGGCTAGT AAAACATGTA AGGAAAATTT TAGGGATGTT 121 AAAGAAAAAA ATAACACAAA ACAAAATATA AAAAAAATCT AACCTCAAGT CAAGGCTTTT 181 CTATGGAATA AGGAATGGAC AGCAGGGGGC TGTTTCATAT ACTGATGACC TCTTTATAGC 241 CAACCTTTGT TCATGGCAGC CAGCATATGG GCATATGTTG CCAAACTCTA AACCAAATAC 301 TCATTCTGAT GTTTTAAATG ATTTGCCCTC CCATATGTCC TTCCGAGTGA GAGACACAAA 361 AAATTCCAAC ACACTATTGC AATGAAAATA AATTTCCTTT ATTAGCCAGA AGTCAGATGC 421 TCAAGGGGCT TCATGATGTC CCCATAATTT TTGGCAGAGG GAAAAAGATC TCAGTGGTAT 481 TTGTGAGCCA GGGCATTGGC CACACCAGCC ACCACCTTCT GATAGGCAGC CTGCACCTGA 541 GGAGTGAATT CTCATTACTT GTACAGCTCG TCCATGCCGA GAGTGATCCC GGCGGCGGTC 601 ACGAACTCCA GCAGGACCAT GTGATCGCGC TTCTCGTTGG GGTCTTTGCT CAGGGCGGAC 661 TGGTAGCTCA GGTAGTGGTT GTCGGGCAGC AGCACGGGGC CGTCGCCGAT GGGGGTGTTC 721 TGCTGGTAGT GGTCGGCGAG CTGCACGCCG CCGTCCTCGA TGTTGTGGCG GATCTTGAAN 781 TTGGCCTTGA TGCCGTTCTT CTGCTTGTCG GCGGTGATAT AGACGTTGTG GCTGTTGTAG 841 TTGTACTCCA GCTTGTGCCN CANGANGTTG CCGTCCTCCT TGAAGTCGAT GCCCTTCANC 901 TCNATGCGGN TCACCNNGGT GTCGCCCNCG AACTTCNCCT CGGCGCGGGN CTNGTANTTG 961 NCGNCNNCCN NGNNGAANNN GGNGNNNNNC NNNNNNGTAN CCNTCNGNNN TNNNNGNNNN 1021 TNNNNNNNNN N //
Primer: SV40pro_F_V2 (Pr0154) Region: SV40 pA,ori Sequence file: RDB16750_A8Jni.seq
>D05919A2_A8Jn_2_SV40pro_F_V2_G02_20_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNNNNGCTT TTGNNNNNGC TAACTTGTTT ATTGCAGCTT 61 ATAATGGTTA CAANNTNNNN GCAATAGCAT CACAAATTTC ACAAATAAAG CATTTTTTTC 121 ACTGCATTCT AGTTGTGGTT TGTCCAAACT CATCAATGTA TCTTATCATG TCTGGATCCG 181 CTGCATTAAT GAATCGGCCA ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC 241 GCTTCCTCGC TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT 301 CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG 361 TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC 421 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA 481 AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT 541 CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG 601 GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG 661 CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT 721 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC 781 AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACNNNNT TCTTGAAGTG GTGGCCTAAC 841 TACNGCTACA CTANANNAAC AGTATTTGGT ATCTGCGCTC TGCTGAANCC NGTTACNTTC 901 GGAAAAAGAN TTGGTAGCTC TTGATNCNNC AAACAAACCA NNNCNGGTAN CGGNGGNTTT 961 TTTTTGTTTG NNNGCAGCNG ATTANNCGCA NAAAAAAAAN GNNCNNNNNN NNNN //
Primer: EGFP-M_R (Pr0743) Region: insert mid(Venus 5',ChRGR(ER) 3') Sequence file: RDB16750_A8Jnj.seq
>16750_16750_B1JcF_EGFP-M_R_E03_15_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN GGNNCTTGNN GTTGGCCTTG ATGCCGTTCT TCTGCTTGTC 61 GGCGGTGATA TAGACGTTGT GGCTGTTGTA GTTGTACTCC AGCTTGTGCC CCAGGATGTT 121 GCCGTCCTCC TTGAAGTCGA TGCCCTTCAG CTCGATGCGG TTCACCAGGG TGTCGCCCTC 181 GAACTTCACC TCGGCGCGGG TCTTGTAGTT GCCGTCGTCC TTGAAGAAGA TGGTGCGCTC 241 CTGGACGTAG CCTTCGGGCA TGGCGGACTT GAAGAAGTCG TGCTGCTTCA TGTGGTCGGG 301 GTAGCGGGCG AAGCACTGCA GGCCGTAGCC CAGGGTGGTC ACGAGGGTGG GCCAGGGCAC 361 GGGCAGCTTG CCGGTGGTGC AGATCAGCTT CAGGGTCAGC TTGCCGTAGG TGGCATCGCC 421 CTCGCCCTCG CCGGACACGC TGAACTTGTG GCCGTTTACG TCGCCGTCCA GCTCGACCAG 481 GATGGGCACC ACCCCGGTGA ACAGCTCCTC GCCCTTGCTC ACCATGGTGG CGACCGGTAT 541 GATCAAACCT TGTATGATAG CCAAAAGGAT GACAATCACG AAGAAGAAGA NNGTGATGTC 601 NAAGATGANG CGGTAGATCT CATACTCGTC TCCTGCTGGG TCTTCANTTT CATCACCNAN 661 GCCCCCTCCN GCACGGNNNC CCACATACAT GTGGNNNATG NNNNACNNNA GCATGNNNNN 721 NNNNN //

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original	Asano, T., Organelle Optogenetics: Direct Manipulation of Intracellular Ca2+ Dynamics by Light. Front. Neurosci. 12: 561 (2018). PMID 30174581. [link to RRC of NBRP]