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pCAG-Arl13bCerulean-Fucci2a

Mammalian expression construct of Arl13bCerulean-Fucci2a

Catalog number RDB16059
Resource name pCAG-Arl13bCerulean-Fucci2a
Clone info. Mammalian expression construct containng the CAG promoter drving Arl13bCerulean-Fucci2a a multicistronic version of Fucci2 composed of Arl13bCerulean, mVenus-hGem(1/110) and mCherry-hCdt1(30/120) fused with t2a and p2a.
This clone was used in subcloning pCDNA5-Frt-CAG-Arl13bCerulean-Fucci2a by the depositor.
Vector backbone pCAGiP (Plasmid)
Selectable markers Ampicillin
Gene/insert name Human CDT1 cDNA Human Gem cDNA Mouse Arl13b cDNA Discosoma sp. RFP cDNA Aequorea victoria GFP cDNA
Depositor|Developer Jackson, Ian | Mort, Richard |
 
Other clones in our bank human CDT1 (NCBI Gene 81620) | human GMNN (NCBI Gene 51053) | mouse Arl13b (NCBI Gene 68146) |

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 agrees to use the BIOLOGICAL RESOURCE only for publication of research papers and the BIOLOGICAL RESOURCE may not be used on humans.
2) In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the literature designated by the DEPOSITOR (Ford, M.J. et al., Dev. Cell 47 (4): 509-523.e5 (2018)) is requested.
3) In publishing the research results to be obtained by use of the BIOLOGICAL RESOURCE, an acknowledgment to the DEPOSITOR and Dr. Richard Mort of Lancaster University is requested.
4) The RECIPIENT must contact the DEPOSITOR in the case of application for any patents or commercial use based on the results from the use of the BIOLOGICAL RESOURCE.
5) The BIOLOGICAL RESOURCE may not be used for commercial purposes by the RECIPIENT.
6) The RECIPIENT may not transfer the BIOLOGICAL RESOURCE to any third parties.
Additional terms and conditions for distribution RECIPIENT is requested to obtain written consent of Dr. Miyawaki, Team of the RIKEN CBS regarding Venus.
Ordering Please visit Information of Request for Distribution.[link] Please contact us. UCSD's MTA is also required.
Order form 
[Credit Card Payment]  [Bank Transfer or Check Payment]
Material Transfer Agreement (MTA) [Word]
Schedule A (GFP-A) [Link]
Approval form [Word] by Dr. Atsushi Miyawaki
Remarks
Please contact us. UCSD's MTA is also required.
Please use Schedule A (GFP-A) additionally.
Please obtain written approval of Dr. Atsushi Miyawaki.
Distribution information (domestic users) [open/close]


Catalog # Resource name Shipping form Fee (non-profit org.)
RDB16059 pCAG-Arl13bCerulean-Fucci2a DNA solution

Ordering Information [in Japanese] [in English]
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 Ford, M.J., A Cell/Cilia Cycle Biosensor for Single-Cell Kinetics Reveals Persistence of Cilia after G1/S Transition Is a General Property in Cells and Mice. Dev Cell. 2018 Nov 19;47(4):509-523.e5. (2018). PMID 30458140.

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

Featured content

Featured content Fucci-related materials (en) (English text)
Featured content Fucci-related materials (ja) (Japanese 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 RDB16059_A9K8p1-1.pdf check

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

Primer: pCAG-F_V2
Region: CAG pro, insert 5' (Arl13b 5')
Sequence file: RDB16059_A9K8a.seq check
>D05769A4_A9K8_1_pCAG-F_V2_C05_08_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNTG NGCGGNNCCG ANTCTGGGAN GNGCCGCCGC ACCCCCTCTA
   61 GCGGGCGCGG GGCGAAGCGG TGCGGCGCCG GCAGGAAGGA AATGGGCGGG GAGGGCCTTC
  121 GTGCGTCGCC GCGCCGCCGT CCCCTTCTCC CTCTCCAGCC TCGGGGCTGT CCGCGGGGGG
  181 ACGGCTGCCT TCGGGGGGGA CGGGGCAGGG CGGGGTTCGG CTTCTGGCGT GTGACCGGCG
  241 GCTCTAGAGC CTCTGCTAAC CATGTTCATG CCTTCTTCTT TTTCCTACAG CTCCTGGGCA
  301 ACGTGCTGGT TATTGTGCTG TCTCATCATT TTGGCAAAGA ATTCCTCGAC ACGCGTGCTA
  361 GCGCTACCGG ACTCAGATCT CGAGCTCAAG CTTATGTTCA GTCTGATGGC CAACTGCTGC
  421 AACTTGTTCA AGCGGTGGCG GGAGCCTGTC AGAAAGGTGA CACTTGTGAT GGTCGGCCTT
  481 GATAATGCTG GTAAAACAGC CACAGCAAAG GGAATCCAAG GAGAGCATCC TGAANACGTA
  541 GCTCCCACTG TTGGCTTTTC TAAAATTGAT CTGAGACAAG GAAAGTTCCA AGTTACCATC
  601 TTTGACTTAN GAGGTGGAAA AAGAATTCGG GGAATTTGGA AGAATTATTA TGCTGAATCC
  661 TATGGGGTAA TATTTGTTGT GGATTCCAGT GATGANGAGA GAATGGAAGA AACNAAGGAG
  721 ACAATGTCCG AANTGCTNAG ACACCCGANG ATATCANGAA AGCCTATATT GGTGCTGGNC
  781 AACAAGCAAG ACNAGGNNGG GGNTTTAGGC GAANCGGANG TGATTGANTG NCTCTCGCTG
  841 GANAANCTGG NCNACGAGCA CAANNGNCTG TGTCNNATCN AANCTTGTTC ANCNNTCTTG
  901 GGANATGGAA ANAAAANTGA NNAGNNNNTN NAAAAGGGGC TTTNNTGGNN NNCTNNNNNT
961 CNTNGNNANN NNNTTTNNNN NCNTNNANNN NNNNCNTCCN NAAAGANNNN NNNNNN
//
Primer: mCerulean-N_R
Region: insert mid (mCerulean, Arl13b 3')
Sequence file: RDB16059_A9K8b.seq check
>D05769A4_A9K8_1_mCerulean-N_R_A06_03_ABI24.ab1
    1 NNNNNNNNNN NNGNNNNNNN NNNNGGNNNC TNGCCGGNGG NGCANATGAN CTTCAGGGTC
   61 AGCTTGCCGT ANGTGGCATC GCCCTCGCCC TCGCCGGACA CGCTGAACTT GTGGCCGTTT
  121 ACGTCGCCGT CCAGCTCGAC CAGGATGGGC ACCACCCCGG TGAACAGCTC CTCGCCCTTG
  181 CTCACCATGG TGGCGACCGG TGGATCCCCT GAGATCGTGT CCTGAGCATC ACCGTTAGGT
  241 CTCTGTCGCA CAGCCAGGGG AGGCAGCGGC TTCCCATAGA AATCATTGTG ACGTGTTTCA
  301 CCAAGAGGCT CAAGTTTTGG AAGTCTAGTG ACTTTGGGGG TTCCCCAGCC AACAGGAGGG
  361 GGAGGTTGGG GAGGCGTGGG ACTCTTTGGA GTAGACTCGT CTGTATTGAC TGGTTCTACC
  421 CGATGACTCC TTTTCATTCT TAGTTTCTTG GTTTTCTTCT TACTGTTTTC CCCGGACTCT
  481 GACCCTCGCT GGTCCTGCTC ATCCTCACTG TCCAGCTGCT GCGACAGTGC CTCCCTGTAA
  541 CTGTCCACCA CTCGCTCGTC AGGGTTTTGG AGGCAGCAAT CTGCCTCACT CTGAGGCGCT
  601 GCTTGCTCTG GCTCCACTTT ATGTTCTAGG ANANCAACAT CACTGTCCTT CTCCACGGTC
  661 TGCTTCTTTT TCTCTTTTTC CTGCTTCTTT TCATTCTCAA TGATTACAGC TGCAATGGGT
  721 TGGANAGGAN TCGCAAGAAC TGGTCCTGAN TCAATCTCAG CCAGACCACN NGTCCCATCA
  781 AGTNNCGTCT GCTCTCGTTC TCTTTCTTCC CTTANNNTCC NGANTCGTTC ANCCCNCTCA
  841 CGTTTCTCTN GTNCCNCNNG ANCTCGCNGT TCANTTNNGN NNTTNNGGAT NCNNNCANTT
901 NNNNNATCNA NNNNCNTNNN NNNNNNN
//
Primer: BGH_rev2
Region: BGH p(A), insert 3' (Gem1_1-110, mVenus)
Sequence file: RDB16059_A9K8c.seq check
>D05769A4_A9K8_1_BGH_rev2_B06_06_ABI24.ab1
    1 NNNNNNNNNN NNNNNGNNNN GGGGNNGGNN NNNNACAGAT GGCTGGCAAC TAGAAGGCAC
   61 AGTCGAGGCT GATCAGCGAG CTCTAGATCA TCGAAGGCGC GCCACATGTG GTACCTTACA
  121 GCGCCTTTCT CCGTTTTTCT GCCACTTCCT TCCAATACTG AGAGGATGGA TTTTCTTTAA
  181 TCATAAGATC AAATGACTCC TGGGTGACTC CTCCAAGATT TTTATTTTCA CTAGATTCTG
  241 GGACAATAAC CCCAGGGCTG GAAGTTGTAG ATGTTAAGTG GTCATTCCGA TGTTTCCTTT
  301 TGGACAAGCC TGCGGACAGC TCATTTTCTC TTCCAACAAG AGATCCAGAT GCAGAAGGCT
  361 GAATCATCTT CAGAGTTCTT CTTGGGACAG AACTATTCTT TATATTCTCT TTGATTTCTT
  421 CTTGTTTCTG CTTCATACTG GGATTCATCT CGAGCGGCCG CCAGTGTGAT GGATATCCCT
  481 TGTACAGCTC GTCCATGCCG AGAGTGATCC CGGCGGCGGT CACGAACTCC AGCAGGACCA
  541 TGTGATCGCG CTTCTCGTTG GGGTCTTTGC TCAGCTTGGA CTGGTAGCTC AGGTAGTGGT
  601 TGTCGGGCAG CAGCACGGGG CCGTCGCCGA TGGGGGTGTT CTGCTGGTAG TGGTCGGCGA
  661 GCTGCACGCC GCCGTCCTCG ATGTTGTGGC GGATCTTGAA GTTGGCCTTG ATGCCGTTCT
  721 TCTGCTTGTC GGCGGTGATA TAGACGTTGT GGCTGTTGTA GTTGTACTCC AGCTTGTGCC
  781 CCANGATGTT GCCGTCCTCC TTGAANTCGA TGCCCTTCAG CTCGATGCGG TTCANCAGGG
  841 NGTCGCCCNC GAACTTCACC TCNGCGCGGG NNTTGTANTT GCCGNCNNCC TTNNNGAANA
901 TGGNGCNCTC CNNGNNNNNN NNNN
//
Primer: P2A_R2
Region: insert mid (mCerulean)
Sequence file: RDB16059_A9K8d.seq check
>D05769A4_A9K8_1_P2A_R2_E05_14_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNTN NGTAGCTCCG CTTCCCTTGT ACNGCTCGTC CNTGCCGAGA
   61 GTGATCCCGN NNNNNNNNNN NAACTCCAGC AGGACCATGT GATCGCGCTT CTCGNTGGNG
  121 TCTTTGCTCA GCTTGGACTG GGTGCTCAGG TAGTGGTTGT CGGGCAGCAG CACGGGGCCG
  181 TCGCCGATGG GGGTGTTCTG CTGGTAGTGG TCGGCGAGCT GCACGCTGCC GTCCTCGATG
  241 TTGTGGCGGA TCTTGAAGTT GGCCTTGATG CCGTTCTTCT GCTTGTCGGC GGTGATATAG
  301 ACGTTGTCGC TGATGGCGTT GTACTCCAGC TTGTGCCCCA GGATGTTGCC GTCCTCCTTG
  361 AAGTCGATGC CCTTCAGCTC GATGCGGTTC ACCAGGGTGT CGCCCTCGAA CTTCACCTCG
  421 GCGCGGGTCT TGTAGTTGCC GTCGTCCTTG AAGAAGATGG TGCGCTCCTG GACGTAGCCT
  481 TCGGGCATGG CGGACTTGAA GAAGTCGTGC TGCTTCATGT GGTCGGGGTA GCGGGCGAAG
  541 CACTGCACGC CCCAGGTCAG GGTGGTCACG AGGGTGGGCC AGGGCACGGG CAGCTTGCCG
  601 GTGGTGCAGA TGAACTTCAG GGTCAGCTTG CCGTAGGTGG CATCGCCCTC GCCCTCGCCG
  661 GACACGCTGA ACTTGTGGCC GTTTACGTCG CCGTCCAGCT CGACCAGGAT GGGCACCACC
  721 CCGGTGAACA GCTCCTCGCC CTTGCTCACC ATGGTGNCGA NCGGTGGATC CCCTGAGATC
  781 GTGTCCTGAG CATCACCGTT AGGTCTCTGT CGCACAGCCA GGGGNNGCAG CGGCTTCCCA
  841 TAGAAATCAT TGTGACGTGT TTCACCAAGA GGCTCAAGTT TTGGAAGTCT ANTGANTTTG
  901 GGGGGTTCCC NAGCCAACAG GNNGGGGGAN GTNGGGGANN NGNGGGNNTC TTTGGANTAN
  961 ANTCGNCNGT ANTGANNNGN NNCNNCCCNA TNNNTNCTTT TCNTTCNTNN NTTNNNNNNN
1021 TTNNNNNNNN NN
//
Primer: mCherry-C_R2
Region: insert mid (mCherry, P2A, mCerulean)
Sequence file: RDB16059_A9K8e.seq check
>D05769A4_A9K8_1_mCherry-C_R2_D05_11_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNT TNNGCTTNNG CCTCTGCTTG ATCTCGCCCT TCNGGGCGCC
   61 GTCCTCGGGG TACATCCGCT CGGAGGAGGC CTCCCAGCCC ATGGTCTTCT TCTGCATTAC
  121 GGGGCCGTCG GAGGGGAAGT TGGTGCCGCG CAGCTTCACC TTGTAGATGA ACTCGCCGTC
  181 CTGCAGGGAG GAGTCCTGGG TCACGGTCAC CACGCCGCCG TCCTCGAAGT TCATCACGCG
  241 CTCCCACTTG AAGCCCTCGG GGAAGGACAG CTTCAAGTAG TCGGGGATGT CGGCGGGGTG
  301 CTTCACGTAG GCCTTGGAGC CGTACATGAA CTGAGGGGAC AGGATGTCCC AGGCGAAGGG
  361 CAGGGGGCCA CCCTTGGTCA CCTTCAGCTT GGCGGTCTGG GTGCCCTCGT AGGGGCGGCC
  421 CTCGCCCTCG CCCTCGATCT CGAACTCGTG GCCGTTCACG GAGCCCTCCA TGTGCACCTT
  481 GAAGCGCATG AACTCCTTGA TGATGGCCAT GTTATCCTCC TCGCCCTTGC TCACCATACG
  541 CGCAGGTCCA GGGTTCTCCT CCACGTCTCC AGCCTGCTTC AGCAGGCTGA AGTTAGTAGC
  601 TCCGCTTCCC TTGTACAGCT CGTCCATGCC GAGAGTGATC CCGGCGGCGG TCACGAACTC
  661 CAGCAGGACC ATGTGATCGC GCTTCTCGTT GGGGTCTTTG CTCAGCTTGG ACTGGGTGCT
  721 CAGGTAGTGG TTGTCGGGCA GCAGCACGGG GCCGTCGCCG ATGGGGGTGT TCTGCTGGTA
  781 GTGGTCGGCG AGCTGCACGC TGCCGTCCTC GATGTTGTGG CGGATCTTGA AGTTGGNCTT
  841 GATGCCGTTC TTCTGCTTGN CGGCGGTGAT ATAGACGTTG TCGCTGATGG CGTTGTANTC
  901 CNGCTNGTGN CCCNNNATGT TGCCGNCCNN CTTGAAGTCN ANGCCCTNCA NNNCGATGCN
961 GNNNCACNNG GGNNNCNNCN NNNAACTTCN NNNNNGCNNN GNNNNNNNAN NNNNNNNNN
//
Primer: mCherry-M_F1
Region: insert mid (mCherry, Cdt1_30-120, T2A, mVenus)
Sequence file: RDB16059_A9K8f.seq check
>D05769A4_A9K8_1_mCherry-M_F1_B05_05_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNA TGNNNAANNN NNNNTGGGCT GGGNNGCCTC CTCCGAGCGG
   61 ATGTACCCCG AGGNNGGCGC CCTGAAGGGC GAGATCAAGC AGAGGCTGAA GCTGAAGGAC
  121 GGCGGCCACT ACGACGCTGA GGTCAAGACC ACCTACAAGG CCAAGAAGCC CGTGCAGCTG
  181 CCCGGCGCCT ACAACGTCAA CATCAAGTTG GACATCACCT CCCACAACGA GGACTACACC
  241 ATCGTGGAAC AGTACGAACG CGCCGAGGGC CGCCACTCCA CCGGCGGCAT GGACGAGCTG
  301 TACAAGGGAT ATCCATCACA CTGGCGGCCG CTCGAGCCCA GCCCCGCCAG GCCCGCACTC
  361 CGCGCCCCGG CCTCCGCTAC CAGTGGCAGC CGCAAGCGCG CCCGCCCGCC CGCCGCCCCC
  421 GGACGCGACC AGGCCAGGCC ACCGGCCCGC AGGAGACTGC GGCTGTCGGT GGACGAGGTT
  481 TCCAGCCCCA GTACCCCCGA GGCCCCAGAC ATCCCAGCCT GCCCTTCTCC GGGCCAGAAG
  541 ATAAAGAAAT CCACCCCGGC AGCAGGTCAG CCGCCCCACC TGACATCCGC GCAGGACCAG
  601 GACACCATCC AATTGGAAGG CCGTGGCTCT CTTCTCACTT GTGGCGATGT CGAAGAGAAT
  661 CCTGGACCCA TGGTGAGCAA GGGCGAGGAG CTGTTCACCG GGGTGGTGCC CATCCTGGTC
  721 GAGCTGGACG GCGACGTAAA CGGCCACAAG TTCAGCGTGT CCGGCGAGGG CGANGGCGAT
  781 GCCACCTACN GCAAGCTGAN CCTNAANCTG ATCTGCACCA CCGGCAAGCT GCCCGNGCCC
  841 TGGCCCACCC TCGNGACCAC CCTGGGCTAC NGNCTGCAGN GNTTCNCCCG CNANCCCNAC
  901 CACNTGAAGC ANCACGACTN CTTCAAGTNC NNCNNGNCCN NNNNNNCNNN CAGNANCGCA
  961 NCATCTNCTN CNNNNANGAN NNNNNNTANN NNANCCNNNN CNANNNNNNN NTNNNNNGNN
1021 NNNNNCNNNN NNNNANNNNN NNNNN
//
Primer: Cdt1_F1
Region: insert mid (T2A, mVenus)
Sequence file: RDB16059_A9K8g.seq check
>D05769A4_A9K8_1_Cdt1_F1_F06_18_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNGNNNNGG NNNNCNTCCN ATTGGAAGGC CGTGGCTCTC
   61 TTCTCNCTTG TGGCGATGTC GAAGAGAATC CTGGACCCAT GGTGAGCAAG GGCGAGGAGC
  121 TGTTCACCGG GGTGGTGCCC ATCCTGGTCG AGCTGGACGG CGACGTAAAC GGCCACAAGT
  181 TCAGCGTGTC CGGCGAGGGC GAGGGCGATG CCACCTACGG CAAGCTGACC CTGAAGCTGA
  241 TCTGCACCAC CGGCAAGCTG CCCGTGCCCT GGCCCACCCT CGTGACCACC CTGGGCTACG
  301 GCCTGCAGTG CTTCGCCCGC TACCCCGACC ACATGAAGCA GCACGACTTC TTCAAGTCCG
  361 CCATGCCCGA AGGCTACGTC CAGGAGCGCA CCATCTTCTT CAAGGACGAC GGCAACTACA
  421 AGACCCGCGC CGAGGTGAAG TTCGAGGGCG ACACCCTGGT GAACCGCATC GAGCTGAAGG
  481 GCATCGACTT CAAGGAGGAC GGCAACATCC TGGGGCACAA GCTGGAGTAC AACTACAACA
  541 GCCACAACGT CTATATCACC GCCGACAAGC AGAAGAACGG CATCAAGGCC AACTTCAAGA
  601 TCCGCCACAA CATCGAGGAC GGCGGCGTGC AGCTCGCCGA CCACTACCAG CAGAACACCC
  661 CCATCGGCGA CGGCCCCGTG CTGCTGCCCG ACAACCACTA CCTGAGCTAC CAGTCCAAGC
  721 TGANCAAAGA CCCCAACGAG AAGCGCGATC ACATGGTCCT GCTGGAGTTC GTGANCGCCG
  781 CCGGGNTCAC TCTCGGCATG NNCGANCTGT ACAANGNNTA TCCATCACAC TGGCGGCCGC
  841 TCNANATGAA TCCNANTATG AAGCANAANN NAAGAAGAAA TCAAAGANNN TATAAAGNNN
901 ANTTCNGNCC NNANNANANN TCNNNANNAN NNNTNNNNNN NNNN
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Ford, M.J., A Cell/Cilia Cycle Biosensor for Single-Cell Kinetics Reveals Persistence of Cilia after G1/S Transition Is a General Property in Cells and Mice. Dev Cell. 2018 Nov 19;47(4):509-523.e5. (2018). PMID 30458140.

2020.10.02

GNP_filter3_RDBDEP_html_200316.pl