Resource data sheet
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pAAV2 SynTetOff Venus-Akaluc

Expression vector of Akaluc luciferase.

Catalog number RDB15782
Resource name pAAV2 SynTetOff Venus-Akaluc
Clone info. Expression vector of Akaluc luciferase.
This clone was constructed based on the pENTR1A-SV40LpA-tTAad-SYN-insulator-TRE-GFP-BGHpA vector (Sohn, J. et al., PLoS One. 12(1): e0169611, 2017).
The following 2 commercial compounds have been demonstrated to be well catalyzed by Akaluc in vivo:
(1) TokeOni (AkaLumine-HCl), Sigma-Aldrich. Product No.: 808350.
(2) AkaLumine-HCl, Wako Pure Chemical Industry. Product No.: 012-26701, 018-26703.
Patent application PCT/JP2017/030491 submitted by RIKEN that covers the creation and use of Akaluc.
Vector backbone pAAV2-DEST(r) (Plasmid)
Gene/insert name firefly (Photinus pyralis) luciferase cDNA
Reference of nucleotide sequence LC320664 (DDBJ accession number)
Depositor Miyawaki, Atsushi |

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 1. The RECIPIENT should obtain written approval from the DEPOSITOR using the APPROVAL FORM prior to entering the AGREEMENT with the RIKEN BRC.
2. A RECIPIENT who belongs to a non-profit organization may use the BIOLOGICAL RESOURCE for an academic research that is non-commercial.
3. In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR or an acknowledgment to the DEPOSITOR are requested.
Iwano, S. et al., Single-cell bioluminescence imaging of deep tissue in freely moving animals. Science 359 (6378): 935-939 (2018).
4. The RECIPIENT agrees to provide a copy of the publication to the DEPOSITOR, oral or written, reporting use of the BIOLOGICAL RESOURCE.
5. The Recipient's Scientist agrees not to transfer the BIOLOGICAL RESOURCE to any third party including any resource centers without the prior written consent of DEPOSITOR.
Additional terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation for the pENTR1A-SV40LpA-tTAad-SYN-insulator-TRE-GFP-BGHpA vector (Sohn, J. et al., PLoS One. 12(1): e0169611, 2017) and an acknowledgment to Dr. Hiroyuki Hioki of the Juntendo University are requested.
The RECIPIENT agrees to use the BIOLOGICAL RESOURCE only for academic research in the non-profit organization.
Distribution information (domestic users) [open/close]


Catalog # Resource name Availability
RDB15782 pAAV2 SynTetOff Venus-Akaluc This clone has been withdrawn.

Ordering Information [in Japanese] [in English]

References and tips

Electronic file

Featured content

Featured content Dr. Miyawaki's resources (English text)

References

original Iwano, S., Single-cell bioluminescence imaging of deep tissue in freely moving animals. Science 359 (6378): 935-939 (2018). PMID 29472486.
original Miyawaki, A., Modified luciferase. Patent publication WO 2018/038248 A1.

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 RDB15782_A8B8p1.pdf

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

Primer: CSII_CMV5_v2 RDB15782_A8B8a.seq
>D04498K4_A8B8_1_CSII_CMV5_V2_A01_01_ABI08.ab1
    1 TGGTGGGAAA TCGGAGCTCG GTACCGCCAC CATGGTGAGC AAGGGCGAGG AGCTGTTCAC
   61 CGGGGTGGTG CCCATCCTGG TCGAGCTGGA CGGCGACGTA AACGGCCACA AGTTCAGCGT
  121 GTCCGGCGAG GGCGAGGGCG ATGCCACCTA CGGCAAGCTG ACCCTGAAGC TGATCTGCAC
  181 CACCGGCAAG CTGCCCGTGC CCTGGCCCAC CCTCGTGACC ACCCTGGGCT ACGGCCTGCA
  241 GTGCTTCGCC CGCTACCCCG ACCACATGAA GCAGCACGAC TTCTTCAAGT CCGCCATGCC
  301 CGAAGGCTAC GTCCAGGAGC GCACCATCTT CTTCAAGGAC GACGGCAACT ACAAGACCCG
  361 CGCCGAGGTG AAGTTCGAGG GCGACACCCT GGTGAACCGC ATCGAGCTGA AGGGCATCGA
  421 CTTCAAGGAG GACGGCAACA TCCTGGGGCA CAAGCTGGAG TACAACTACA ACAGCCACAA
  481 CGTCTATATC ACCGCCGACA AGCAGAAGAA CGGCATCAAG GCCAACTTCA AGATCCGCCA
  541 CAACATCGAG GACGGCGGCG TGCAGCTCGC CGACCACTAC CAGCAGAACA CCCCCATCGG
  601 CGACGGCCCC GTGCTGCTGC CCGACAACCA CTACCTGAGC TACCAGTCCG CCCTGAGCAA
  661 AGACCCCAAC GAGAAGCGCG ATCACATGGT CCTGCTGGAG TTCGTGACCG CCGCCGGGAT
  721 CACTCTCGGC ATGGACGAGC TGTACAAGGG ATCCATGGAA GATGCCAAAA ACATTAAGAA
  781 GGGCCCAGCG CCGTTCTACC CACTCGAAGA CGGGACCGCC GGCGAGCAGC TGCACAAAGC
  841 CATGAAGCGC TACGCCCTGG TGCCCGGCGC CATCGCCTTT ACCGACGCAC ATATTCAGGT
  901 GGACGTTACC TACGCCGAGT ACTTCGAGAT GAGCGTTCGG CTGGCAGAAG CTATGAGGCG
  961 CTATGGGCTG AATACAAACC ATCGGATCGT GGTGTGCAGC GAGAATAGCT CGCAGTTCTT
 1021 TCATGCCCGT GTTGGGTGCC CTGTTCATCG GTGTGGCTGG TGACCCAGGC TAACGACATC
1081 TACACGAGCG CGAGCCTGGC TGGCACCAGC AATGGGGTCA TCAAGTCCAA GGCGAC
//
Primer: Akaluc 1215-1243 RDB15782_A8B8b.seq
>D04498K4_A8B8_1_Akaluc 1215-1243_D01_04_ABI08.ab1
    1 TCCTGCGGGC GCTCGCCTAC TGGGACGAGG ACGAGCACTT CTTCATCGTG GACCGGCTGA
   61 AGAGCCTGAT CAAATACAAG GGCTACCAGG TAGCCCCAGC CGAACTGGAG GGCATCCTGC
  121 TGCAACACCC CTACATCTTC GACGCCGGAG TCGCCGGCCT GCCCGACGAC GATGCCGGCG
  181 AGCTGCCCGC CGCAGTCGTC GTGTTGGAAC ACGGTAAAAC CATGACCGAG AAAGAGATCG
  241 TGGACTATGT GGCCAGCCAG GTTACAACCG CCAAGAAGCT GCGCGGTGGT GTTGTGTTTG
  301 TGGATGAAGT CCCTAGAGGA TCGACCGGCA AGTTAGACGC CCGCAAGATC CGCGAGATTC
  361 TCACTAAGGC CAAGAAGGAC GGCAAGATCG CCGTGTAAAC GCGTCTGTGC CTTCTAGTTG
  421 CCAGCCATCT GTTGTTTGCC CCTCCCCCGT GCCTTCCTTG ACCCTGGAAG GTGCCACTCC
  481 CACTGTCCTT TCCTAATAAA ATGAGGAAAT TGCATCGCAT TGTCTGAGTA GGTGTCATTC
  541 TATTCTGGGG GGTGGGGTGG GGCAGGACAG CAAGGGGGAG GATTGGGAAG ACAATAGCAG
  601 GCATGCTGGG GATGCGGTGG GCTCTATGGG CGGCCGCGTC GAGATATCTA GACCCAGCTT
  661 TCTTGTACAA AGTGGTACGC GTGCGGCCGC AGGAACCCCT AGTGATGGAG TTGGCCACTC
721 CCTCTCTGCG CGCT
//
Primer: Venus-C_F2 RDB15782_A8B8c.seq
>D04498K4_A8B8_1_Venus-C_F2_B01_02_ABI08.ab1
    1 TGCATTGGAC GGCGCAGGAA TCACTCTCGG CATGGACGAG CTGTACAAGG GATCCATGGA
   61 AGATGCCAAA AACATTAAGA AGGGCCCAGC GCCGTTCTAC CCACTCGAAG ACGGGACCGC
  121 CGGCGAGCAG CTGCACAAAG CCATGAAGCG CTACGCCCTG GTGCCCGGCG CCATCGCCTT
  181 TACCGACGCA CATATTCAGG TGGACGTTAC CTACGCCGAG TACTTCGAGA TGAGCGTTCG
  241 GCTGGCAGAA GCTATGAGGC GCTATGGGCT GAATACAAAC CATCGGATCG TGGTGTGCAG
  301 CGAGAATAGC TCGCAGTTCT TCATGCCCGT GTTGGGTGCC CTGTTCATCG GTGTGGCTGT
  361 GGCCCCAGCT AACGACATCT ACAACGAGCG CGAGCTGCTG AACAGCATGG GCATCAGCCA
  421 GCCCACCGTC GTATTCGTGA GCAAGAAAGG GCTGCGAAAG GTCCTCAACG TGCAAAAGAA
  481 GCTACCGATC ATACGAAAGA TCATCATCAT GGATAGCAAG ACCGACTACC AGGGCTTCCA
  541 AAGCATGTAC ACCTTCGTGA CTTCCCATTT GCCACCCAGC TTCAACGAGT ACGACTTCGT
  601 GCCCGAGAGC TTCGACCGGG ACAAAACCAT CGCCCTGATC ATGAACAGTA GTGGTAGTAC
  661 AGGATTACCC AAGGGCGTAG CCCTACCGCA CCGCACCGCT TGTGTCCGAT TCAGTCATGC
  721 CCGCGACCCC ATCTTCGGCT ACCAGAACAT CCCCGACACC GCTATCCTCA GCGTGGTGCC
  781 ATTTCACCAC GGCTTCGGCA TGTTCACCAC GCTGGGCTAC TTGATCTGCG GCTTTCGGGT
  841 CGTGCTCATG TACCGCTTCG AGGAGGAGCT ATTCTTGCGC AGCTTGCAAG ACTATAAGAT
  901 TCAATCTGCC CTGCTGGTGC CCACACTATT TAGCTGCCTC GCTAAGAGCA CTCTCATCGA
  961 CAAGTACGAC CTAAAGCAGC TTGCGCGAGA TCGCCCAGCG GGCGGGGGCG CCCGCTCAGC
 1021 AAGAGGGTAG GTGAGGGCCG GTGGACCAAC GCTTCCGCCC TACCAGCAAT CCGCCAGGGC
 1081 TATGGCCCTG ACAGGAACAC CAACGGTCGG TCCATGATTC ACCCCCCCGG AAGGGGGGGA
1141 ACCGGT
//
Primer: Akaluc 576-593 RDB15782_A8B8d.seq
>D04498K4_A8B8_1_Akaluc 576-593_C01_03_ABI08.ab1
    1 CAAAAAAACC AGGGGGCGTA GCCCTACCGC ACCGCACCGC TTGTGTCCGA TTCAGTCATG
   61 CCCGCGACCC CATCTTCGGC TACCAGAACA TCCCCGACAC CGCTATCCTC AGCGTGGTGC
  121 CATTTCACCA CGGCTTCGGC ATGTTCACCA CGCTGGGCTA CTTGATCTGC GGCTTTCGGG
  181 TCGTGCTCAT GTACCGCTTC GAGGAGGAGC TATTCTTGCG CAGCTTGCAA GACTATAAGA
  241 TTCAATCTGC CCTGCTGGTG CCCACACTAT TTAGCTGCCT CGCTAAGAGC ACTCTCATCG
  301 ACAAGTACGA CCTAAGCAGC TTGCGCGAGA TCGCCAGCGG CGGGGCGCCG CTCAGCAAGG
  361 AGGTAGGTGA GGCCGTGGCC AAACGCTTCC GCCTACCAGG CATCCGCCAG GGCTATGGCC
  421 TGACAGAAAC AACCAACGCC GTCATGATCA CCCCCGAGGG GGACCGTAAG CCTGGCTCAG
  481 TAGGCAAGGT GGTGCCCTTC TTCGAGGCTA AGGTGGTAGA CTTGGTCACC GGTAAGACAC
  541 TGGGTGTGAA CCAGCGCGGT GAGCTGTGCG TCCGTGGCCC CATGATCATG AGCGGCTACG
  601 TTAACAACCC CGAGGCTACG AACGCTCTCA TCGACAAGGA CGGCTGGCTG CACAGCGGCG
  661 ACATCGCCTA CTGGGACGAG GACGAGCACT TCTTCATCGT GGACCGGCTG AAGAGCCTGA
  721 TCAAATACAA GGGCTACCAG GTAGCCCCAG CCGAACTGGA GGGCATCCTG CTGCAACACC
  781 CCTACATCTT CGACGCCGGA GTCGCCGGCC TGCCCGACGA CGATGCCGGC GAGCTGCCCG
  841 CCGCAGTCGT CGTGTTGGAA CACGGTAAAA CCATGACCGA GAAAGAGATC GTGGACTATG
  901 TGGCCAGCCA GGTTACAACC GCCAAGAAGC TGCGCGGTGG TGTTGTGTTT GTGGATGAAG
  961 TCCCTAGAGG GATCGACCGG CAAGTTAGAC GCCCGCAAGG ATCCGCGAGA TTCTCACTAA
 1021 AGGCCCAAGA AGGACGGCAG ATCGCCCGTG TAAACGCGGT CTGTGCCTTC TAGCTGCAGT
 1081 CATCTATTGA TTGCCCGACC CCGATGGCAT TCGTTGGACC CTGGGAAGGG TGGCCCACAT
1141 C
//
Primer: tTA-Ad R RDB15782_A8B8e.seq
>D04498K4_A8B8_1_tTA-AdR_E01_05_ABI08.ab1
    1 GATCCGGGGG GCAGGAGGTT TATGGACTTT GCTCTTGTCC AGTCTAGACA TGGTGAATTC
   61 GATATCAAGC TTATCGATAC CGTCCGCCGC AGCGCAGATG GTCGCGCCCG TGCCCCCCTA
  121 TCTCGCGCCT CGCGTGGTGC GGTCCGGCTG GGCCGGCGGC GGCGCGGACG CGACCAAGGT
  181 GGCCGGGAAG GGGAGTTTGC GGGGGACCGG CGAGTGACGT CAGCGCGCCT TCAGTGCTGA
  241 GGCGGCGGTG GCGCGCGCCG CCAGGCGGGG GCGAAGGCAC TGTCCGCGGT GCTGAAGCTG
  301 GCAGTGCGCA CGCGCCTCGC CGCATCCTGT TTCCCCTCCC CCTCTCTGAT AGGGGATGCG
  361 CAATTTGGGG AATGGGGGTT GGGTGCTTGT CCAGTGGGTC GGGGTCGGTC GTCAGGTAGG
  421 CACCCCCACC CCGCCTCATC CTGGTCCTAA AACCCACTTG CACTCATACG CAGGGCCCTC
  481 TGCAGCTCGA CCGCGCCGCC CCGGGCGGCT CGCTGCTGCC CCCTAGCGGG GGAGGGACGT
  541 AATTACATCC CTGGGGGCTT TGGGGGGGCT CGAGTTTACT CCCTATCAGT GATAGAGAAC
  601 GTATGTCGAG TTTACTCCCT ATCAGTGATA GAGAACGATG TCGAGTTTAC TCCCTATCAG
  661 TGATAGAGAA CGTATGTCGA GTTTACTCCC TATCAGTGAT AGAGAACGTA TGTCGAGTTT
  721 ACTCCCTATC AGTGATAGAG AACGTATGTC GAGTTTATCC CTATCAGTGA TAGAGAACGT
  781 ATGTCGAGTT TACTCCCTAT CAGTGATAGA GAACGTATGT CGAGGTAGGC GTGTACGGTG
  841 GGAGGCCTAT ATAAGCAGAG CTCGTTTAGT GAACCGTCAG ATCGCCTGGA GAATTCGAGC
  901 TCGGTACCGC CACCATGGTG AGCAAAGGGC GAGGAGCTGT TCACCGGGGT GGTGCCCATC
  961 CTGGTCGAGC TGGACGGGCG ACGTAAACGG GCCACAAGTT TCAGCGTGTC CGGGCGAGGG
 1021 CGAGGCGATG CCACCTACGG CAAGCTGACC CTGAAGCTGA TCTGCACCAC CGGGCCAGCT
 1081 GCCGTGCCCT GACCATCTTC GTGACAACCC TGGGCCTACC GACCCTGCCA GTGCCTTTCG
1141 AACCCCGC
//
Primer: tTA-Ad F RDB15782_A8B8f.seq
>D04498K4_A8B8_1_tTA-AdF_F01_06_ABI08.ab1
    1 ACATTCTTTT CGAGCTGGAA CTAATCATAT GTGGCCTGGA GAAACAGCTA AAGTGCGAAA
   61 GCGGCGGGCC GGCCGACGCC CTTGACGATT TTGACTTAGA CATGCTCCCA GCCGATGCCC
  121 TTGACGACTT TGACCTTGAT ATGCTGCCTG CTGACGCTCT TGACGATTTT GACCTTGACA
  181 TGCTCCCCGG GTAACTAAGT AAGGATCTCA GACATGATAA GATACATTGA TGAGTTTGGA
  241 CAAACCACAA CTAGAATGCA GTGAAAAAAA TGCTTTATTT GTGAAATTTG TGATGCTATT
  301 GCTTTATTTG TAACCATTAT AAGCTGCAAT AAACAAGTTA ACAACAACAA TTGCATTCAT
  361 TTTATGTTTC AGGTTCAGGG GGAGGTGTGG GAGGTTTTTT AAAGCAAGTA AAACCTCTAC
  421 AAATGTGGTA AGATCCAAAG CCTGCTTTTT TGTACAAACT TGTCACGTGC GGACCGAGCG
481 GCCGCAGGAA CCCCTAGTGA TGGAGTTGGC CACTCCCTCT CTGCGCGCT
//

Please visit Sequencing and PCR primers for primer information.


References

References

original Iwano, S., Single-cell bioluminescence imaging of deep tissue in freely moving animals. Science 359 (6378): 935-939 (2018). PMID 29472486.
original Miyawaki, A., Modified luciferase. Patent publication WO 2018/038248 A1.
reference Nasu, Y., Unnaturally aglow with a bright inner light. Science 359 (6378): 868-869 (2018). PMID 29472468.
reference Kuchimaru, T., A luciferin analogue generating near-infrared bioluminescence achieves highly sensitive deep-tissue imaging. Nat. Commun. 7: 11856 (2016). PMID 27297211.
reference Iwano, S., Development of simple firefly luciferin analogs emitting blue, green, red, and near-infrared biological window light. Tetrahedron 69 (19): 3847-3856 (2013).
reference Iwano, S., Development of simple firefly luciferin analogs emitting blue, green, red, and near-infrared biological window light. Tetrahedron 69 (19): 3847-3856 (2013).

2019.03.20

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