Resource data sheet
Prev.

pMRX-IP-GFP-LC3-RFP-LC3 delta G

Fluorescent probe to evaluate autophagic flux by the rat LC3 fused with GFP and RFP.

Catalog number RDB14600
Resource name pMRX-IP-GFP-LC3-RFP-LC3 delta G
Clone info. Fluorescent probe to evaluate autophagic flux. Expression vector of rat microtubule-associated protein 1 light chain 3 beta (LC3) fused with GFP and RFP. This probe is cleaved by endogenous ATG4 proteases into equimolar amounts of GFP-LC3 and RFP-LC3deltaG. The GFP-LC3 is degraded by autophagy, while RFP-LC3deltaG remains in the cytosol as an internal control. Thus, autophagic flux can be estimated by calculating the GFP/RFP signal ratio. Please refer DISCUSSION section of Kaizuka, T., Mol. Cell 64, 835-849, 2016 for the difference between pMRX-IP-GFP-LC3-RFP and this clone.
Expression was confirmed by the depositor with immunoblotting and fluorescent microscopy. Please refer Saitoh, T. et al., FEBS Lett., 532 (1-2): 45-51, (2002) for the construction of pMRX-IP vector.
Vector backbone pMRX-IP (Plasmid)
Selectable markers Amp^r (E. coli), Puro^r (mammalian cells).
Gene/insert name Rat LC3 cDNA Aequorea victoria GFP cDNA Discosoma sp. RFP cDNA
Depositor Mizushima, Noboru |
 
Other clones in our bank Rat Map1lc3b (KEGG orthology K10435) |

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
Ordering Information [in Japanese] [in English]
Terms and conditions set forth by the DEPOSITOR (1) BIOLOGICAL RESOURCE is limited to for academic research in non-profit organization. (2) The RECIPIENT of BIOLOGICAL RESOURCE shall obtain a prior written consent on use of it from the DEPOSITOR. (3) Any use of the BIOLOGICAL RESOURCE for profit purposes by a non-profit organization or any use of the BIOLOGICAL RESOURCE by a profit organization requires a separate license from the DEPOSITOR prior to distribution. (4) In publishing research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the original publication designated by the DEPOSITOR (Kaizuka, T. et al. Mol. Cell, 64 (4): 835-849, 2016) and an acknowledgment to the DEPOSITOR are requested. (5) The RECIPIENT must contact the DEPOSITOR in case of application for any patents or commercial use with the results from the use of BIOLOGICAL RESOURCE.
(Japanese text [open/close])
Additional terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOUECE, the BIOLOGICAL RESOUECE should be described as pMRX-IP-GFP-LC3-RFP-LC3 delta G, which has the pMX backbone.

(Japanese text [open/close])
Remarks ((Additional form)) Approval Form(FormD), GFP-A
((Remarks)) Remember that you will be working with samples containing infectious virus.Please contact us. We will provide you with appropriate documents.
(Japanese text [open/close])

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB14600 pMRX-IP-GFP-LC3-RFP-LC3 delta G DNA solution

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 RDB14600_A7Bap1_2.pdf

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

Primer: EGFP-C RDB14600_A7Baa.seq
>D04280A1_A7Ba_1_EGFP-C_A04_01_ABI24.ab1
    1 ACGGGGGCAC ATCTCGGCAT GGACGAGCTG TACAGGGTGG CGGTGGCTCG GGCGGTGGTG
   61 GGTCGGGTGG CGGCGGATCT ATCGATGAAT TGAATTCCAT GCCGTCCGAG AAGACCTTCA
  121 AACAGCGCCG GAGCTTCGAA CAAAGAGTGG AAGATGTCCG GCTCATCCGG GAGCAGCACC
  181 CCACCAAGAT CCCAGTGATT ATAGAGCGAT ACAAGGGTGA GAAGCAGCTG CCCGTCCTGG
  241 ACAAGACCAA GTTCCTTGTA CCTGATCACG TGAATATGAG CGAACTCATC AAGATAATTA
  301 GAAGGCGCCT GCAGCTCAAT GCTAACCAAG CCTTCTTCCT CCTGGTGAAT GGGCACAGCA
  361 TGGTGAGTGT GTCCACACCC ATCTCTGAAG TGTACGAGAG CGAGAGAGAT GAAGACGGCT
  421 TCCTGTACAT GGTCTATGCC TCCCAGGAGA CGTTCGGGAT GGCCTCCTCC GAGGACGTCA
  481 TCAAGGAGTT CATGCGCTTC AAGGTGCGCA TGGAGGGCTC CGTGAACGGC CACGAGTTCG
  541 AGATCGAGGG CGAGGGCGAG GGCCGCCCCT ACGAGGGCAC CCAGACCGCC AAGCTGAAGG
  601 TGACCAAGGG CGGCCCCCTG CCCTTCGCCT GGGACATCCT GTCCCCTCAG TTCCAGTACG
  661 GCTCCAAGGC CTACGTGAAG CACCCCGCCG ACATCCCCGA CTACTTGAAG CTGTCCTTCC
  721 CCGAGGGCTT CAAGTGGGAG CGCGTGATGA ACTTCGAGGA CGGCGGCGTG GTGACCGTGA
  781 CCCAGGACTC CTCCCTGCAG GACGGCGAGT TCATCTACAA GTGAAGCTGC GCGGCACCAA
  841 CTTCCCCTCC GACGGCCCCG TAATGCAGAA GAGACCATGG GCTGGGAGGC CTCCACGAGC
  901 CGGATGTTAC CCCGAGGACG GCGCCCTGAG GCGAGATCAG ATGAGCTGAA GCTGAAGGAC
  961 GGCGGCACTA CGACGCGAGG TCAGACCACT ACATGGCCAG AGCCGGTGCA GCTGCCCGGC
 1021 GCTACAAGAA CGGACTCAAG CTGGAACATC ACCTCCCACA ACGAAGACTA CACCAATCGT
 1081 GAACAGTACG AGGCGGCGAG GGCGCCACTC ACGGCATCAT GAATTGAATC ATGGCTCGGA
1141 AGACTCAACG GCCGACTGAA CAAGTGATAA GTTCCGTCAT CTGGCAC
//
Primer: mRFP1-inner1 RDB14600_A7Bab.seq
>D04280A1_A7Ba_1_mRFP1-inner1_E10_13_ABI24.ab1
    1 GCGCGAGGGG GGGGCTGACG TGACCCAGGA CTCCTCCCTG CAGGACGGCG AGTTCATCTA
   61 CAAGGTGAAG CTGCGCGGCA CCAACTTCCC CTCCGACGGC CCCGTAATGC AGAAGAAGAC
  121 CATGGGCTGG GAGGCCTCCA CCGAGCGGAT GTACCCCGAG GACGGCGCCC TGAAGGGCGA
  181 GATCAAGATG AGGCTGAAGC TGAAGGACGG CGGCCACTAC GACGCCGAGG TCAAGACCAC
  241 CTACATGGCC AAGAAGCCCG TGCAGCTGCC CGGCGCCTAC AAGACCGACA TCAAGCTGGA
  301 CATCACCTCC CACAACGAGG ACTACACCAT CGTGGAACAG TACGAGCGCG CCGAGGGCCG
  361 CCACTCCACC GGCGCCATCG ATGAATTGAA TTCCATGCCG TCCGAGAAGA CCTTCAAACA
  421 GCGCCGGAGC TTCGAACAAA GAGTGGAAGA TGTCCGGCTC ATCCGGGAGC AGCACCCCAC
  481 CAAGATCCCA GTGATTATAG AGCGATACAA GGGTGAGAAG CAGCTGCCCG TCCTGGACAA
  541 GACCAAGTTC CTTGTACCTG ATCACGTGAA TATGAGCGAA CTCATCAAGA TAATTAGAAG
  601 GCGCCTGCAG CTCAATGCTA ACCAAGCCTT CTTCCTCCTG GTGAATGGGC ACAGCATGGT
  661 GAGTGTGTCC ACACCCATCT CTGAAGTGTA CGAGAGCGAG AGAGATGAAG ACGGCTTCCT
  721 GTACATGGTC TATGCCTCCC AGGAGACGTT CTAAGGATCC CAGTGTGGTG GTACGGGAAT
  781 TCCTGCAGGC CTCGAGGGCC GGCGCGCCGC GGCCGCTACG TAAATTCCGC CCCCCCCCCC
841 CCTCTCCCTC
//
Primer: MMLV-pol_F2 RDB14600_A7Bac.seq
>D04280A1_A7Ba_1_MMLV-pol_F2_C04_07_ABI24.ab1
    1 TACGGGGGGG GTGGACATCC TCTAGACTGC CGGATCTCCC GCCGCCACCA TGGTGAGCAA
   61 GGGCGAGGAG CTGTTCACCG GGGTGGTGCC CATCCTGGTC GAGCTGGACG GCGACGTAAA
  121 CGGCCACAAG TTCAGCGTGT CCGGCGAGGG CGAGGGCGAT GCCACCTACG GCAAGCTGAC
  181 CCTGAAGTTC ATCTGCACCA CCGGCAAGCT GCCCGTGCCC TGGCCCACCC TCGTGACCAC
  241 CCTGACCTAC GGCGTGCAGT GCTTCAGCCG CTACCCCGAC CACATGAAGC AGCACGACTT
  301 CTTCAAGTCC GCCATGCCCG AAGGCTACGT CCAGGAGCGC ACCATCTTCT TCAAGGACGA
  361 CGGCAACTAC AAGACCCGCG CCGAGGTGAA GTTCGAGGGC GACACCCTGG TGAACCGCAT
  421 CGAGCTGAAG GGCATCGACT TCAAGGAGGA CGGCAACATC CTGGGGCACA AGCTGGAGTA
  481 CAACTACAAC AGCCACAACG TCTATATCAT GGCCGACAAG CAGAAGAACG GCATCAAGGT
  541 GAACTTCAAG ATCCGCCACA ACATCGAGGA CGGCAGCGTG CAGCTCGCCG ACCACTACCA
  601 GCAGAACACC CCCATCGGCG ACGGCCCCGT GCTGCTGCCC GACAACCACT ACCTGAGCAC
  661 CCAGTCCGCC CTGAGCAAAG ACCCCAACGA GAAGCGCGAT CACATGGTCC TGCTGGAGTT
  721 CGTGACCGCC GCCGGGATCA CTCTCGGCAT GGACGAGCTG TACAAGGGTG GCGGTGGCTC
  781 GGGCGGTGGT GGGTCGGGTG GCGGCGGATC TATCGATGAA TTGAATTCCA TGCCGTCCGA
  841 GAAGACCTTC AAACAGCGCC GGAGCTTCGA ACAAAGAGTG GAAGATGTCC GGCTCATCCG
  901 GGAGCAGCAC CCCACCAAGA TCCCAGTGAT TATAGAGCGA TACAAGGGTG AGAAGCAGCT
  961 GCCCGTCCTG ACAGACCAAG TTCCTTGTAC CTGATCACGT GAATATGAGC GAACTCATCA
 1021 AGATAATTTA GAAGCGCTGC AGCTCAATGC TAACCAAGCC TTCTTCTCTG CTGATGGGCA
 1081 CAGCATGTGA GTGTGTCCAC ACCCATCTCT GAGTGTACGA AGCGAGAGAT GAGACGCTCC
1141 TGTACATGTC TATGCTCCAG GAGAAACGTT CCGG
//
Primer: EGFP-N RDB14600_A7Bad.seq
>D04280A1_A7Ba_1_EGFP-N_E04_13_ABI24.ab1
    1 GAATCAACCC GGGTTTAAAG CTCCTCGCCC TTGCTCACCA TGGTGGCGGC GGGAGATCCG
   61 GCAGTCTAGA GGATGGTCCA CCCCCGGGGT CGGCAGCCTT CACGTGGGCG GCGTGTATCC
  121 AAGCTGCGAT GCCGTCTACT TTGAGGGCGG TGGGGGTGGT CAGCAGGACT GTGTAAGGTC
  181 CTTTCCAGCG AGGTTCTAGG TTCTTAGTCT GGTGTCGGCG GACCCACACT GTGTCGCCGA
  241 CTCGGTAAGG GTGAGGTACC ACCGGTCGGT CCAGTTGTTC TTGGTAGGCT GCCGCCAGAG
  301 GTCTCCAGAC TTCGTGCTGG ACTAAGTAGA GAGCCTGTAA GTGAGCTTGG AGAGAGGGGC
  361 TGTTAGTAAC TCTTGTCATG TCAGGGTCAG GGAAGTTTAC AAGGGGCGGG GGTGCCCCAT
  421 ATAAGATCTC ATATGGCCAT ATGGGGGCGC CTAGAGAAGG AGTGAGGGCT GGATAAAGGG
  481 AGGATCGAGG CGGGGTCGAA CGAGGAGGTT CAAGGGGGAG AGACGGGGCG GATGGAGGAA
  541 GAGGAGGCGG AGGCTTAGGG TGTACAAAGG GCTTGACCCA GGGAGGGGGG TCAAAAGCCA
  601 AGGCTTCCCA GGTCACGATG TAGGGGACCT GGTCTGGGTG TCCATGCGGG CCAGGTGAAA
  661 AGACCTTGAT CTTAACCTGG GTGATGAGGT CTCGGTTAAA GGTGCCGTCT CGCGGCCATC
  721 CGACGTTAAA GGTTGGCCAT TCTGCAGAGC AGAAGGTAAC CCAACGTCTC TTCTTGACAT
  781 CTACCGACTG GTTGTGAGCG ATCCGCTCGA CATCTTTCCA GTGACCTAAG GTCAAACTTA
  841 AGGGAGTGGT AACAGTCTGG CCCGGGCCCA TATTTTCAGA CAAATACAGA AACACAGTCA
  901 GACAGAGACA ACACAGAACG ATGCTGCAGC AGACAAGACG CGCGGCGCGG CTTCGGTCCC
  961 AAACCGAAAG CAAAAAATTC AGACGGAGGC GGGAACTGTT TTAAGTTCTC GTCTCCTACC
 1021 AGGAACCAAC AATTATCCCT CCTCTAAGGG GGGGTGCACC CAAAGAAAGT CCAAAACCGA
 1081 TTCGGGAATT TTTGGAACCT CAAGTCGGGG CCACAAAAAC GGCCCTCGAA GTCCCTGGGA
 1141 ACGTCTCAGA TGCCGCGGGG TTTCGAACTC GTTCAGGTTC CCACCAACCG GGGGTCGCGC
1201 CCCCGGATAT
//
Primer: pJB8-Forward RDB14600_A7Bae.seq
>D04280A1_A7Ba_1_pJB8-Forward_D04_10_ABI24.ab1
    1 GTCCGTTCCC ATTGCATACG TTGTATCCAT ATCATAATAT GTACATTTAT ATTGGCTCAT
   61 GTCCAACATT ACCGCCATGT TGACATTGAT TATTGACTAG TTATTAATAG TAATCAATTA
  121 CGGGGTCATT AGTTCATAGC CCATATATGG AGTTCCGCGT TACATAACTT ACGGTAAATG
  181 GCCCGCCTGG CTGACCGCCC AACGACCCCC GCCCATTGAC GTCAATAATG ACGTATGTTC
  241 CCATAGTAAC GCCAATAGGG ACTTTCCATT GACGTCAATG GGTGGAGTAT TTACGGTAAA
  301 CTGCCCACTT GGCAGTACAT CAAGTGTATC ATATGCCAAG TACGCCCCCT ATTGACGTCA
  361 ATGACGGTAA ATGGCCCGCC TGGCATTATG CCCAGTACAT GACCTTATGG GACTTTCCTA
  421 CTTGGCAGTA CATCTACGTA TTAGTCATCG CTATTACCAT GCTAGAGAAC CATCAGATGT
  481 TTCCAGGGTG CCCCAAGGAC CTGAAATGAC CCTGTGCCTT ATTTGAACTA ACCAATCAGT
  541 TCGCTTCTCG CTTCTGTTCG CGCGCTTCTG CTCCCCGAGC TCAATAAAAG AGCCCACAAC
  601 CCCTCACTCG GCGCGCCAGT CCTCCGATTG ACTGAGTCGC CCGGGTACCC GTGTATCCAA
  661 TAAACCCTCT TGCAGTTGCA TCCGACTTGT GGTCTCGCTG TTCCTTGGGA GGGTCTCCTC
  721 TGAGTGATTG ACTACCCGTC AGCGGGGGTC TTTCATTTGG GGGCTCGTCC GGGATCGGGA
  781 GACCCCTGCC CAGGGACCAC CGACCCACCA CCGGGAGGTA AGCTGGCCAG CAACTTATCT
  841 GTGTCTGTCC GATTGTCTAG TGTCTATGAC TGATTTTATG CGCCTGCGTC GGTACTAGTT
  901 AGCTAACTAG CTCTGTATCT GGCGGACCCG TGGTGGAACT GACGAGTTCG GAACACCCGG
  961 CCGCAACCCT GGGAGACGTC CCAGGACTTC GGGGGCCGTT TTTGTGGCCC GACCTGAGTC
 1021 CAAAAATCCC GATCGTTTGG ACTCTTTGGT GCACCCCCCC TTTAGAGAGG GATATGTGGT
 1081 CTGTAGGAGA CGAGATCTAA ACAGTCCCGC TCGTCTGAAT TTGCCTCGGT TGAACCGAGC
1141 CGCGCGCGCG TCTTCTCTGC CTGCCACGCA AATCCGC
//
Primer: p15Aori_F RDB14600_A7Baf.seq
>D04280A1_A7Ba_1_p15Aori_F_F04_16_ABI24.ab1
    1 GCAGGGACGT GATTTTTGTG ATGCTCGTCA GGGGGGCGGA GCCTATGGAA AAACGCCAGC
   61 AACGCGGCCT TTTTACGGTT CCTGGCCTTT TGCTGGCCTT TTGCTCACAT GTTCTTTCCT
  121 GCGTTATCCC CTGATTCTGT GGATAACCGT ATTACCGCCT TTGAGTGAGC TGATACCGCT
  181 CGCCGCAGCC GAACGACCGA GCGCAGCGAG TCAGTGAGCG AGGAAGCGGA AGAGCGCCCA
  241 ATACGCAAAC CGCCTCTCCC CGCGCGTTGG CCGATTCATT AATGCAACTA TGGCCATTTA
  301 ATGTAAATAC TTAAGAAAAA AAACCAAATT AATTTTGATA CATGCTGCAT GTGAAAGACC
  361 CCCGCTGACG GGTAGTCAAT CACTCAGAGG AGACCCTCCC AAGGAACAGC GAGACCACAA
  421 GTCGGATGCA ACTGCAAGAG GGTTTATTGG ATACACGGGT ACCCGGGCGA CTCAGTCAAT
  481 CGGAGGACTG GCGCCCCGAG TGAGGGGTTG TGGGCTCTTT TATTGAGCTC GGGGAGCAGA
  541 AGCGCGCGAA CAGAAGCGAG AAGCGAACTG ATTGGTTAGT TCAAATAAGG CACAGGGTCA
  601 TTTCAGGTCC TTGGGGCACC CTGGAAACAT CTGATGGTTC TCTAGAAACT GCTGAGGGCT
  661 GGACCGCATC TGGGGACCAT CTGTTCTTGG CCCTGAGCCG GGGCAGGAAC TGCTTACCAC
  721 AGATATCCTG TTTGGCCCAT ATTCAGCTGT TCCATCTGTT CTTGGCCCTG AGCCGGGGCA
  781 GGAACTGCTT ACCACAGATA TCCTGTTTGG CCCATATTCA GCTGTTCCAT CTGTTCCTGA
  841 CCTTGATCTG AACTTCTCTA TTCTCAGTTA TGTATTTTTC CATGCCTTGC AAAATGGCGT
  901 TACTTAAGCT AGCTTGCCAA ACCTACAGGT GGGGTCTTTC ATTCCCCCCT TTTTCTGGAG
  961 ACTAAATAAA ATCTTTTATT TTATCGTCGA GCCATGGGGT CGTGCGCTCC TTTCGGTCGG
 1021 GCGCTGCGGG TCGTGGGGCG GGGCGTCAGG CACCGGGCTT GCGGGTCATG CACCAGTGCG
 1081 CGTCCTCCGG CACCTCGACG TCGCGTGACG TGAAGCCGAG TCGCTCGTTA GAGGAGTTGC
1141 GGACGCGGAG TCTCTCCAGG AATGC
//
Primer: pMXs-3LTRup_R2 RDB14600_A7Bag.seq
>D04280A1_A7Ba_1_pMXs-3LTRup_R2_G04_19_ABI24.ab1
    1 GGCATATAAA ATCTTTTATT TTATCGTCGA GCCATGGGGT CGTGCGCTCC TTTCGGTCGG
   61 GCGCTGCGGG TCGTGGGGCG GGCGTCAGGC ACCGGGCTTG CGGGTCATGC ACCAGGTGCG
  121 CGGTCCTTCG GGCACCTCGA CGTCGGCGGT GACGGTGAAG CCGAGCCGCT CGTAGAAGGG
  181 GAGGTTGCGG GGCGCGGAGG TCTCCAGGAA GGCGGGCACC CCGGCGCGCT CGGCCGCCTC
  241 CACTCCGGGG AGCACGACGG CGCTGCCCAG ACCCTTGCCC TGGTGGTCGG GCGAGACGCC
  301 GACGGTGGCC AGGAACCACG CGGGCTCCTT GGGCCGGTGC GGCGCCAGGA GGCCTTCCAT
  361 CTGTTGCTGC GCGGCCAGCC GGGAACCGCT CAACTCGGCC ATGCGCGGGC CGATCTCGGC
  421 GAACACCGCC CCCGCTTCGA CGCTCTCCGG CGTGGTCCAG ACCGCCACCG CGGCGCCGTC
  481 GTCCGCGACC CACACCTTGC CGATGTCGAG CCCGACGCGC GTGAGGAAGA GTTCTTGCAG
  541 CTCGGTGACC CGCTCGATGT GGCGGTCCGG GTCGACGGTG TGGCGCGTGG CGGGGTAGTC
  601 GGCGAACGCG GCGGCGAGGG TGCGTACGGC CCGGGGGACG TCGTCGCGGG TGGCGAGGCG
  661 CACCGTGGGC TTGTACTCGG TCATGGTTGT GGCCATATTA TCATCGTGTT TTTCAAAGGA
  721 AAACCACGTC CCCGTGGTTC GGGGGGCCTA GACGTTTTTT TAACCTCGAC TAAACACATG
  781 TAAAGCATGT GCACCGAGGC CCCAGATCAG ATCCCATACA ATGGGGTACC TTCTGGGCAT
  841 CCTTCAGCCC CTTGTTGAAT ACGCTTGAGG AGAGCCATTT GACTCTTTCC ACAACTATCC
  901 AACTCACAAC GTGGCACTGG GGTTGTGCCG CCTTTGCAGG TGTATCTTAT ACACGTGGCT
  961 TTTGGGCCGC AGAGGCACCT GTCGCCAGGG TGGGGGGTTC CGCTGCCTGC AAAGGGTCGC
 1021 TACAGACGTT GTTTGTCTTC AGAGCTTCCA GAGGAACTGC TTCTTCACGA CATTCAACAG
 1081 ACCTTGCATT CCTTGCGAGA GGGAAGACCC CTAGGATGGC TCGTCAGAGA CCGTCAGTAT
1141 TCGGCCTCAC ATTGCCTAAA GAAACGGCCA AATATG
//

Please visit plasmidSequencing and PCR primers for primer information.

References and tips

Electronic file

Featured content

Featured content Autophagy (English text)

References

original Kaizuka, T., An Autophagic Flux Probe that Releases an Internal Control. Mol Cell. 2016 Nov 17;64(4):835-849 (2016). PMID 27818143.

2017.08.30

GNP_filter3_RDBDEP_html_170829.pl