RDB15246 - tPA-mK-GO-N1

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tPA-mK-GO-N1 (#RDB15246)

Tissue-type plasminogen activator (tPA)-tagged fluorescent timer protein, monomeric Kusabira Green Orange (mK-GO).

Clone info.	Tissue-type plasminogen activator (tPA)-tagged fluorescent timer protein, monomeric Kusabira Green Orange (mK-GO). Green, yellow, and red fluores-cence of dense-core vesicle in PC12 cells were implicated as new, intermediate, and old ages, respectively (Tsuboi, T., Mol. Biol. Cell 21 (1): 87-94 (2010)).
Comment	This clone contains a cDNA fragment corresponding to the 92nt to 1771nt of NM_013151.3 for rat tPA sequence. Known differences (at least) : G to A substitution at 174nt (Arg28Lys). ; T to C substitution at 607nt (172Asn, synonym). ; A to G substitution at 642nt (Lys184Arg).
Vector backbone	pEGFP-C1 (Plasmid)
Selectable markers	Kanamycin (E. coli), G418 (mammalian cell).
Gene/insert name	Fungia concinna Kusabira Green Orange cDNA Rat tPA cDNA
Depositor\|Developer	Miyawaki, Atsushi \|

Distribution information

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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.
Tsuboi, T. et al., Mol. Biol. Cell 21 (1): 87-94 (2010).

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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. Tsuboi, T. et al., Mol. Biol. Cell 21 (1): 87-94 (2010).

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. Tsuboi, T. et al., Mol. Biol. Cell 21 (1): 87-94 (2010).
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.)
RDB15246	tPA-mK-GO-N1	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 tPA-mK-GO-N1 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15246).

Reference section:

Tsuboi, T., Kitaguchi, T., Karasawa, S., Fukuda, M., Miyawaki, A., Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine cells revealed by newly developed monomeric fluorescent timer protein. Mol. Biol. Cell. 21 (1): 87-94 (2010). PMID 19889833. [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	RDB15246_B1Kpp1-1.pdf

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

Primer: CMV_Forward (Pr0016) Region: CMV pro, insert 5' (tPA 5') Sequence file: RDB15246_B1Kpa.seq
>D04498F4_B1Kp_2_CMV_Forward_A06_01_ABI08.ab1 1 NNNNNNNNNN NNNNNNNNNN NTNNNNNNNN NTGGGCGGTA GGCGTGTACG GTGGGAGGTC 61 TATATAAGCA GAGCTGGTTT AGTGAACCGT CAGATCCGCT AGCGCTACCG GACTCAGATC 121 TCGAGCTCAA GCTTCGAATT CTGCAGTCGA CATGAAGGGA GAGCTGTTGT GCGTCCTGCT 181 GCTTTGTGGA GTGGCGTTCA CGTTGCCTGA CCAGGGAATA CACAGGAGGT TCAAAAGAGG 241 AGCTCGGTCC TACAGAGCGA CCTGCAGAGA TGAACAGACT CAGACAACTT ACCAACAGCA 301 TCAGTCATGG CTACGTCCCA TGCTCAGAGG CAATCGGGTG GAATACTGCC GGTGCAACAG 361 CGGCCTGGCA CAGTGCCACT CGGTGCCCGT CCGAAGTTGC AGTGAACCGA GATGCTTCAA 421 TGGGGGTACG TGTCAGCAGG CCCTGTATTT CTCTGACTTC GTCTGCCAGT GCCCTGACGG 481 ATTTGTTGGG AAACGCTGTG ATATAGATAC CAGAGCAACC TGCTTCGAGG GCCAGGGCAT 541 CACCTACAGA GGCACATGGA GCACAGCAGA AAATGGGGCT GAATGCATCA ACTGGAATAG 601 CAGTGCCCTG TCCCAGAAGC CCTACAGTGC AAGGAGGCCA AATGCCATCA AGCTGGGCCT 661 TGGGAACCAC AATTACTGCA GAAACCCAGA CCGAGACGTG AGGCCCTGGT GCTATGTCTT 721 TAAGGCAGGG AAGTATACCA CGGAGTTCTG CAGCACGCCG GCTTGCCCTA ANGGANCAAC 781 TGAGGACTGC TATGTTGGAA AAGGTGTGAC TTACCGTGGC ACCCACNGCT TTACCACATC 841 CAAGGCCTCC TGCCTCCCAT GGAATTCCAT GATCCTGATA GGNNAGACTT ACACAGCGNG 901 NANGGNCNAC TCCCANGCAC TTGGCCNNNN NAGANNNAAT TATTGCCGGN NNCNNNATGG 961 GGNNGNCAAA CCTTGGNNNC NNNNGATGNN NNNNNAANNC TGANNTGGGA ANATTGCNNN 1021 NTGNCCCNNN NNNNNNN //
Primer: SV40polyA_R2 (Pr0623) Region: insert 3' (mK-GO 3') Sequence file: RDB15246_B1Kpb.seq
>D04498F4_B1Kp_2_SV40polyA_R2_B06_02_ABI08.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNNNNNGNN NNNNNNNNNN ATTGCNTTCA TTTTATGTTT 61 CAGGTTCAGG GGGAGGTGTG GGAGGTTTTT TAAAGCAAGT AAAACCTCTA CAAATGTGGT 121 ATGGCTGATT ATGATCTAGA GTCGCGGCCG CTTAGGAATG AGCTACTGCA TCTTCTACCA 181 GCTCAGTAAT GTTGCCTTCG GTTTTCCTGA CGAGGCGATG GCCGATGTAA TGGTCTCCTG 241 GCATTTCAAG AATCTCTTTT GCCGCCTTGT AAGTAGTCTT GAATTGGCAT TTGTGATTGC 301 CGCCTCCTTC AAGTTTTAGG TACATCGTAA CATCACCCTT CAGAACTCCG TCGCTGGCAG 361 TAATTTTCTC GGTTGATGGC TCCCAATCAA CACTTTGGTT TTGCATGATA GGACCATCGG 421 CAGGAAAGTT AACCCCAGTA AATTTGGATT TGTGGTAGAA GGTGTTTCCT CTAAGGCTTA 481 TATGCGCACT GACTGAAGCG GACCCACCAT CTTCGAACTC CAACGACCTT TCCCATGACA 541 GGCCTTCAGG AAATGCTTGT TTGAAATAGT CTGGTATCTC TTCTGGATAT TTAGTAAATA 601 CTCTGTGGCC GTAACAGAAC ACGTGTGACA CTAAGTCAAA CGCGAAAGGC ATTGGCCCGC 661 CCTCGGCCAT TGTGACGCGT AGTGTCATCT CTTGATGTCC CTCGTAAGGT CTGCCTGTGC 721 CTTCACCTTC AATTGTGAAC TCATGCCCAT TGACGGAGCC GTCCATGTAG TACCTCATCT 781 TCATCTCTGG TTTANTCACA CTCNCCATGA NCGGTGGATC CCGGGNCCGC GGTTGCTTCA 841 TGTTGTCTTG NATCCANTTC ANGTAATTAG TGACCTTTGT GTATATCCCT GGCACGNCCT 901 TNTGCCCACA GCCNANGNCC CAGCTGATGA TGNCCANNNA AGTCATNCGN NTNNCGATCA 961 TGCNCANNNN NNGGNCNCCT GANNNANCCT GGCANNCGTC ANNNNNGTCN TNGNNTNNCN 1021 NCNNNNNNCN GGNNNNNCTN CTGNNNNNNN NNN //
Primer: mKO2-N_R (Pr0494) Region: insert mid (mK-GO 5', tPA 3') Sequence file: RDB15246_B1Kpc.seq
>D04498F4_B1Kp_2_mKO2-N_R_D06_04_ABI08.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNG NNNNNNNNGT CNNTGTNNNN NNTCATCTTC 61 ATCTCTGGTT TAATCACACT CACCATGACC GGTGGATCCC GGGCCCGCGG TTGCTTCATG 121 TTGTCTTGGA TCCAGTTCAG GTAATTAGTG ACCTTTGTGT ATATCCCTGG CACGTCCTTC 181 TGCCCACAGC CGAGGCCCCA GCTGATGATG CCCAGTAAAG TCATCCGCTT ATCGATCATG 241 CACACCAGAG GGCCTCCTGA GTCACCCTGG CACGCGTCAT GGACGTCTTG GTTGCCCCCA 301 GTTCGGGTGT CTCCTGCACA CAGCATGTTG CTCGTGATGG TTTTGTTAAA CAGATGCTGT 361 GAGGTACAGC GGCTGGACGG ATACAGTCTG ACGTGAGCCT CCTTCAGCCG GTCAGAGAAG 421 AAAGGAGAGG ATGCCTCATG CTTGCCGTAG CCAGAAAGCT CACACTCTGT CCAGTCAGGA 481 AGCTGTACGT CGGGGTCAGG GAGGCAGGCA GTGCCGACAG AACTGCTCTC CTGGGCACAC 541 TGACTGGAAT CTGACCTCAG CTGCAGTAAT GCGATGTCAT TGTCATAAGT GTCGTCATCA 601 AATTCCTTAT GGACTATGTA CTTTTCGATC TCGAATGTCT GCTCCTCCTC TCCAGGGACC 661 ACTCTGTATG TTCTGCCCAA GACCACTTTA AGATGATGGG GTGGAAACCT NNNNNNNAAA 721 GCAGTGGGCG GCAGATAGCA CCCAGCANGA ACTGATCAGC ACCCCTCNNN NCANGANNCT 781 CTCTNCNNGG ANACCTCTTG TTCNTGACAA AGATGGCGNN CTGCCNNNGN TGTGANGTGA 841 NGTCTGTGAA GAGTCCNNCN TTTAANTNCG AAACNGANGC TGTTTGTATT GNCCTCNNNC 901 CGCNNNGGAN CATGGGGNAC NTGTCGCAAT NNTTNCCATG TNNGCTTTNN GNCNNTTNNN 961 NACGNGGCAC CAAGGTTNNN NNNNNCCCCN TNTGGNNNNC NNNNANNANT TGNGTCTNNC 1021 CNNNNNNANN NNNNNGNNAN NTNGNCCNNC NNCGCNNNNN NANNNNNTNN NNNNNNNN //
Primer: SV40pro_ori_F (Pr0731) Region: SV40 pro_ori, NeoR_KanR Sequence file: RDB15246_B1Kpd.seq
>D04498F4_B1Kp_2_SV40pro_ori_F_C09_03_ABI08.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNCTCGGCC TCTGANCTAT TCCNGAAGTA GTGAGGAGGC 61 TTTTTTGGAG GCCTAGGCTT TTGCAAAGAT CGATCAAGAG ACAGGATGAG GATCGTTTCG 121 CATGATTGAA CAAGATGGAT TGCACGCAGG TTCTCCGGCC GCTTGGGTGG AGAGGCTATT 181 CGGCTATGAC TGGGCACAAC AGACAATCGG CTGCTCTGAT GCCGCCGTGT TCCGGCTGTC 241 AGCGCAGGGG CGCCCGGTTC TTTTTGTCAA GACCGACCTG TCCGGTGCCC TGAATGAACT 301 GCAAGACGAG GCAGCGCGGC TATCGTGGCT GGCCACGACG GGCGTTCCTT GCGCAGCTGT 361 GCTCGACGTT GTCACTGAAG CGGGAAGGGA CTGGCTGCTA TTGGGCGAAG TGCCGGGGCA 421 GGATCTCCTG TCATCTCACC TTGCTCCTGC CGAGAAAGTA TCCATCATGG CTGATGCAAT 481 GCGGCGGCTG CATACGCTTG ATCCGGCTAC CTGCCCATTC GACCACCAAG CGAAACATCG 541 CATCGAGCGA GCACGTACTC GGATGGAAGC CGGTCTTGTC GATCAGGATG ATCTGGACGA 601 AGAGCATCAG GGGCTCGCGC CAGCCGAACT GTTCGCCAGG CTCAAGGCGA GCATGCCCGA 661 CGGCGAGGAT CTCGTCGTGA CCCATGGCGA TGCCTGCTTG CCGAATATCA TGGTGGAAAA 721 TGGCCGCTTT TCTGGATTCA TCGACTGTGG CCGGCTGGGT GTGGCGGANC GCTATCAGGA 781 CATAGCGTTG GCTACCCGTG ATATTGCTGA ANAGCTTGGC GGCGAATGGG CTGANCGCTT 841 CCTCGTGCTT TACGGTATCG CCGCTCCCGA TTCGCANCGC ATCNCCTTCT ATCGCCTTCN 901 TGANGAGTTC TTCTGANCNG GANTCTGGGG GTTCGAAATG ACCGACNNAG CNNCNCCCNA 961 CCNGNCNTCN CNANNTTTCN ANTNNNNNGN CNNCNNTCNA TNNNNNNNGG GCTTCNNNNN 1021 NNNTTNNNNG NNNNNNNNNN //

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original	Tsuboi, T., Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine cells revealed by newly developed monomeric fluorescent timer protein. Mol. Biol. Cell. 21 (1): 87-94 (2010). PMID 19889833. [link to RRC of NBRP]