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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.
Comment 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|Developer Seki, Takahiro |
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External Database
Human GAPDH

          Reference sequence
            

          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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          提供条件 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 (Sato, M. et al. Genes Cells 21 (8): 861-873, 2016)。
          付加的提供条件 本件リソースの使用は学術機関での学術研究に限る。本件リソースの使用にあたって、利用者はプロメガ社の限定使用条件に従う必要がある。
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          RDB15088 pcDNA5-GAPDH-HaloTag DNA solution

          Please review the QC test results indicated by check icon below as well as clone information before placing your order.

          References and tips

          Electronic file

          Original reference

          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.

          Further references such as user reports and related articles (go to bottom)

          Featured content

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

          Sequence information

          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 RDB15088_A7F8p1-3.pdf check

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

          Primer: CMV_Forward
          Region: CMV pro,T7 pro,insert 5'(hGAPDH 5')
          Sequence file: RDB15088_A7F8a.seq check
          >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
          Region: insert mid(HaloTag,TEV site,hGPDH 3')
          Sequence file: RDB15088_A7F8b.seq check
          >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
          Region: bGHpA,insert 3'(HaloTag 3')
          Sequence file: RDB15088_A7F8c.seq check
          >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
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

          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.

          2022.05.17

          GNP_filter3_RDBDEP_html_220516.pl