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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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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).
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提供依頼書  [依頼書の記入例 ]
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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 check

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 check
>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 check
>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 check
>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 check
>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]