Resource data sheet
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pcDNA5-GAPDH-HaloTag

Expression vector for the visualization of chaperone-mediated autophagy (CMA).

Catalog number RDB15088
Resource name pcDNA5-GAPDH-HaloTag
Alternative name GAPDH-HT/pcDNA5/FRT
Clone info. Expression vector of human glyceraldehyde 3-phosphate dehydrogenase (GAPDH), tagged with C-terminal HaloTag, for genomic integration. Translocation of the GAPDH-HT labeled with an appropriate fluorescent dye as a substrate of chaperone-mediated autophagy (CMA) from cytosol to lysosome can be detected. Refer Fig. 1 of Seki, T., et al., PLoS ONE 7 (2), e31232, 2012.
Expression was confirmed by the depositor with immunostaining.
Vector backbone pcDNA5/FRT (Plasmid)
Size of vector backbone 5.1 kb
Selectable markers Amp^r (E. coli), Hyg^r (mammalian cells).
Gene/insert name Human GAPDH cDNA
Depositor Seki, Takahiro |
 
Other clones in our bank human GAPDH (NCBI Gene 2597) |

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested (Sato, M. et al. Genes Cells 21 (8): 861-873, 2016).
Additional terms and conditions for distribution The use of the BIOLOGICAL RESOURCE is restricted to the academic researches conducted by non-profit organization. By using this material, the RECIPIENT agrees to be bound by the conditions of the limited use statement of the Promega Corporation.
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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB15088 pcDNA5-GAPDH-HaloTag DNA solution

Ordering Information [in Japanese] [in English]

References and tips

Electronic file

Featured content

Featured content Dnaconda's recommendation EXR0071e (English text)
Featured content Dnaconda's recommendation EXR0071j (Japanese text)
Featured content Autophagy (English text)

References

original Sato, M., Fluorescent-based evaluation of chaperone-mediated autophagy and microautophagy activities in cultured cells. Genes Cells 21 (8): 861-873 (2016). PMID 27377049.
original Seki,T., Establishment of a novel fluorescence-based method to evaluate chaperone-mediated autophagy in a single neuron. PLoS One 7 (2): e31232 (2012). PMID 22363588.

User reports and related articles (go to bottom)


Sequence information

RIKEN BRC has sequenced portions of this material for quality test. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB15088_A7F8p1.pdf

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

Primer: CMV-Forward RDB15088_A7F8a.seq
>D04515A1_A7F8-1_CMV-Forward_A09_01_ABI08.ab1
    1 ATTTTCACTT CCGCCCCATT GACGCAAATG GGCGGTAGGC GTGTACGGTG GGAGGTCTAT
   61 ATAAGCAGAG CTCTCTGGCT AACTAGAGAA CCCACTGCTT ACTGGCTTAT CGAAATTAAT
  121 ACGACTCACT ATAGGGAGAC CCAAGCTGGC TAGCGTTTAA ACTTAAGCTT GGTACCGAGC
  181 TCGGATCCAC TAGTCCAGTG TGGTGGAATT CTGCAGATAT CACCATGGGG AAGGTGAAGG
  241 TCGGAGTCAA CGGATTTGGT CGTATTGGGC GCCTGGTCAC CAGGGCTGCT TTTAACTCTG
  301 GTAAAGTGGA TATTGTTGCC ATCAATGACC CCTTCATTGA CCTCAACTAC ATGGTTTACA
  361 TGTTCCAATA TGATTCCACC CATGGCAAAT TCCATGGCAC CGTCAAGGCT GAGAACGGGA
  421 AGCTTGTCAT CAATGGAAAT CCCATCACCA TCTTCCAGGA GCGAGATCCC TCCAAAATCA
  481 AGTGGGGCGA TGCTGGCGCT GAGTACGTCG TGGAGTCCAC TGGCGTCTTC ACCACCATGG
  541 AGAAGGCTGG GGCTCATTTG CAGGGGGGAG CCAAAAGGGT CATCATCTCT GCCCCCTCTG
  601 CTGATGCCCC CATGTTCGTC ATGGGTGTGA ACCATGAGAA GTATGACAAC AGCCTCAAGA
  661 TCATCAGCAA TGCCTCCTGC ACCACCAACT GCTTAGCACC CCTGGCCAAG GTCATCCATG
  721 ACAACTTTGG TATCGTGGAA GGACTCATGA CCACAGTCCA TGCCATCACT GCCACCCAGA
  781 AGACTGTGGA TGGCCCCTCC GGGAAACTGT GGCGTGATGG CCGCGGGGCT CTCCAGAACA
  841 TCATCCCTGC CTCTACTGGC GCTGCCAAGG CTGTGGGCAA AGGTCATCCC TGAGCTGAAC
  901 GGGAAGCTCA CTGGCATGGC CTTCCGTGTC CCCACTGCCA ACGTGTCAGT GGGTGGACCT
  961 GACCTGCCGT CTAGAAAACC CTGGCCCAAT ATGATGACAT CAAGAAAGTT GTGGAGCAGC
 1021 GTCGGAGGAC CCCTTCAAGG CATCCTTGGC TACACTGAGC ACAGATGATC TCCTCTGACC
1081 TTC
//
Primer: HaloTag-N_R RDB15088_A7F8b.seq
>D04515A1_A7F8-1_HaloTag-N_R_B09_02_ABI08.ab1
    1 ATAGCATCAT ATGGGGGTCG ATGGAAGCCA GTACCGATTT CGGATCCATC GTTATCGCTC
   61 TGAAAGTACA GATCCTCAGT GGTTGGCTCG AGAGCTCGAG ATCCCTCCTT GGAGGCCATG
  121 TGGGCCATGA GGTCCACCAC CCTGTTGCTG TAGCCAAATT CGTTGTCATA CCAGGAAATG
  181 AGCTTGACAA AGTGGTCGTT GAGGGCAATG CCAGCCCCAG CGTCAAAGGT GGAGGAGTGG
  241 GTGTCGCTGT TGAAGTCAGA GGAGACCACC TGGTGCTCAG TGTAGCCCAG GATGCCCTTG
  301 AGGGGGCCCT CCGACGCCTG CTTCACCACC TTCTTGATGT CATCATATTT GGCAGGTTTT
  361 TCTAGACGGC AGGTCAGGTC CACCACTGAC ACGTTGGCAG TGGGGACACG GAAGGCCATG
  421 CCAGTGAGCT TCCCGTTCAG CTCAGGGATG ACCTTGCCCA CAGCCTTGGC AGCGCCAGTA
  481 GAGGCAGGGA TGATGTTCTG GAGAGCCCCG CGGCCATCAC GCCACAGTTT CCCGGAGGGG
  541 CCATCCACAG TCTTCTGGGT GGCAGTGATG GCATGGACTG TGGTCATGAG TCCTTCCACG
  601 ATACCAAAGT TGTCATGGAT GACCTTGGCC AGGGGTGCTA AGCAGTTGGT GGTGCAGGAG
  661 GCATTGCTGA TGATCTTGAG GCTGTTGTCA TACTTCTCAT GGTTCACACC CATGACGAAC
  721 ATGGGGGCAT CAGCAGAGGG GGCAGAGATG ATGACCCTTT TGGCTCCCCC CTGCAAATGA
  781 GCCCCAGCCT TCTCCATGGT GGTGAAGACG CCAGTGGACT CCACGACGTA CTCAGCGCCA
  841 GCATCGCCCC ACTTGATTTT GGAGGGGATC TCGCTCCTGG AAAGATGGTG ATGGGGATTT
  901 CCATTGATGA CAAGCTTTCC CGTTTCTCAG CCCTTGACGG TGCCATGGGA AATTTGCCCA
  961 TGGGTGGGAA TCATATTTGG GAACATGTAA ACCCATGTAG TTTGAGGTTC CATTGAGGGG
 1021 TCATTGATGG CCACCATATC AACTTACCAG AGTAAAAGGC CAGCCCCTTG GGTTGACCAG
1081 GGACGC
//
Primer: BGH_rev2 RDB15088_A7F8c.seq
>D04515A1_A7F8-1_BGH_rev2_C09_03_ABI08.ab1
    1 CATTAGGGTT CGGGAGGCAC GGGGGAGGGG CAAACAACAG ATGGCTGGCA ACTAGAAGGC
   61 ACAGTCGAGG CTGATCAGCG GGTTTAAACG GGCCCTCTAG ACTCGAGCGG CCGCCCCAAG
  121 GGGTTATGCT AGTTATTGCT CAGCGGTGGC AGCAGCCAAC TCAGCTTCCT TTCGGGCTTT
  181 GTTAGCAGCC GGATCAGCTT GCATGCCTGC AGGTCGACTC TAGAATTCTA TTAACCGGAA
  241 ATCTCCAGAG TAGACAGCCA GCGCGCGATC TCGCTGCCGA TCAGGTCCGG GTTGTCTTCT
  301 TGCAGCAGAT TCAGACCCGG GCCGATGTCC ACAGCCTTGC AGTTAGGCAG GCTTTTGGCC
  361 AGGCGAGCGG CTTCGGCCGG TGGGATCAGA ACGCCTGGGG TGCCCCAGAA CAGCAGCTTC
  421 GGGACAGGGG ACTGGTGCAG CCAGTCCATG TATTCTTCGA CCAGCGCGAC GATGTTCGCT
  481 GGCTCACCGG CGATTGGCAG CTCGTTTGGG AAGCGCCACA GTGGCTCGCG GTCAACAGGA
  541 TTCAGGAACG GCTCGCGGTA ATGGTCCATC TCGACTTCAG TCAGCGGGCG GACGACACCC
  601 ATCGGCAGCG TACCCTCGAT AAAAACGTTC TGATCGATGA TCAGCTTGCG GCCGACGTCG
  661 GTGGTGCGGA AGGCCTGGAA GGTCTCGCGG GCAAATTCTG GCCATTCGTC CCAGGTCGGG
  721 ATAGGGCGGA TGAACTCCAT AAATGCAATA CCTTTGACGC GCTCTGGATT GCGCTTGGCC
  781 CAGTGGAAAC CCAGAGCGGA GCCCCAGTCG TGAATGACCA GGACGACCTC TTCCAGACCC
  841 AGGGCTTCGA TGAAGGCATC CATGAAGCGG ACGTGGTCGT CGAAGAAATA ACCCAGGGTC
  901 TGGTTTGTCG GATTTGCCCA TACCGATCAG GTCTGGGAGC AATGCAGCGA TGGGTCGGTG
  961 CACATGCGGG ATGATGTTGC GCCACACGTA GAGGAGGTCG GGTTACCCGT GCCAGACAGC
 1021 ACAGGGTGCA ATCGCGCGGA ACAACATCGA CGTAGTGCAT GCGCTCGCCA GGAACTTCCG
1081 ACATAA
//
Primer: BGH_pA_F RDB15088_A7F8d.seq
>D04515A1_A7F8-1_BGHpA_F_D09_04_ABI08.ab1
    1 CGTGAAGGTT ATTCTATTCT GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG GGAGGATTGG
   61 GAAGACAATA GCAGGCATGC TGGGGATGCG GTGGGCTCTA TGGCTTCTGA GGCGGAAAGA
  121 ACCAGCTGGG GCTCTAGGGG GTATCCCCAC GCGCCCTGTA GCGGCGCATT AAGCGCGGCG
  181 GGTGTGGTGG TTACGCGCAG CGTGACCGCT ACACTTGCCA GCGCCCTAGC GCCCGCTCCT
  241 TTCGCTTTCT TCCCTTCCTT TCTCGCCACG TTCGCCGGCT TTCCCCGTCA AGCTCTAAAT
  301 CGGGGGCTCC CTTTAGGGTT CCGATTTAGT GCTTTACGGC ACCTCGACCC CAAAAAACTT
  361 GATTAGGGTG ATGGTTCACG TACCTAGAAG TTCCTATTCC GAAGTTCCTA TTCTCTAGAA
  421 AGTATAGGAA CTTCCTTGGC CAAAAAGCCT GAACTCACCG CGACGTCTGT CGAGAAGTTT
  481 CTGATCGAAA AGTTCGACAG CGTCTCCGAC CTGATGCAGC TCTCGGAGGG CGAAGAATCT
  541 CGTGCTTTCA GCTTCGATGT AGGAGGGCGT GGATATGTCC TGCGGGTAAA TAGCTGCGCC
  601 GATGGTTTCT ACAAAGATCG TTATGTTTAT CGGCACTTTG CATCGGCCGC GCTCCCGATT
  661 CCGGAAGTGC TTGACATTGG GGAATTCAGC GAGAGCCTGA CCTATTGCAT CTCCCGCCGT
  721 GCACAGGGTG TCACGTTGCA AGACCTGCCT GAAACCGAAC TGCCCGCTGT TCTGCAGCCG
  781 GTCGCGGAGG CCATGGATGC GATCGCTGCG GCCGATCTTA GCCAGACGAG CGGGTTCGGC
  841 CCATTCGGAC CGCAAGGAAT CGGTCAATAC ACTACATGGC GTGATTTCAT ATGCGCGATT
  901 GCTGATCCCC ATGTGTATCA CTGGGCAAAC TGTGATGGAC GACACCGTCA GTGCGTCCGT
  961 CGCGCAGGCT CTCGATGAGC TGATGCTTTG GCCCGAGGAC TGCCCCGAAA GTCCCGGCAC
 1021 TCGTGCACGC GGGATTTCGG CTCAACCATG TCTGGACGGA CCATTGCGCA TAACAGTCGA
1081 TCAATGGACC TGGAGCGGAG GCGCAATGGA T
//

Please visit Sequencing and PCR primers for primer information.


References

References

original Sato, M., Fluorescent-based evaluation of chaperone-mediated autophagy and microautophagy activities in cultured cells. Genes Cells 21 (8): 861-873 (2016). PMID 27377049.
original Seki,T., Establishment of a novel fluorescence-based method to evaluate chaperone-mediated autophagy in a single neuron. PLoS One 7 (2): e31232 (2012). PMID 22363588.

2018.09.20

GNP_filter3_RDBDEP_html_180823.pl