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

pMGP (#RDB15790)

Expression vector of EGFP and pac under the control of Perkinsus marinus MOE gene promoter.

Clone info.	Bicistronic expression vector of EGFP and pac intervened by a self-cleaving 2A peptide, which is controled under the Perkinsus marinus MOE (PmMOE) promoter. This clone is for a drug selection system using puromycin for transfecting Perkinsus species. Perkinsus is sensitive to puromycin, and 100 microgram/ml of this drug completely blocks its proliferation.
Comment	Expression was confirmed by the depositor with fluorescence microscopy.
Vector backbone	pCR4-TOPO (Plasmid)
Size of vector backbone	4 kb
Selectable markers	Amp^r, Kan^r.
Gene/insert name	Perkinsus marinus MOE genomic DNA Aequorea victoria GFP cDNA Streptomyces alboniger puromycin-N-acetyl-transferase (pac) genomic DNA
Depositor|Developer	Nozaki, Tomoyoshi | Matsuzaki, Motomichi |

 

Sequence	RDB15790z.seq

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 forms	Order form [Credit Card Payment] [Bank Transfer Payment] [Example of order form ] MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ] Please visit Information of Request for Distribution.[link] For for-profit-research purpose, please contact us.
Terms and conditions for distribution	In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of one of the literature designated by the DEPOSITOR is requested (Sakamoto, H. et al., Parasitol. Int. 69: 13-16 (2018); Sakamoto, H. et al., J. Eukaryot. Microbiol. in press).

提供案内 (日本国内) [open/close]

Catalog #	Resource name	Shipping form	Fee (non-profit org.)
RDB15790	pMGP	DNA solution

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

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How to cite this biological resource

Materials & Methods section:

The pMGP was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15790).

Reference section:

Sakamoto, H., Kita, K., Matsuzaki, M., A Novel 2A-peptide-containing Plasmid to Generate Stable Perkinsus marinus Cells Expressing Organelle-targeted Genes. J. Eukaryot. Microbiol. 68 (5): e12861 (2021). PMID 34051022. [link to RRC of NBRP]

Sakamoto, H., Hirakawa, Y., Ishida, K., Keeling, P.J., Kita, K., Matsuzaki, M., Puromycin selection for stable transfectants of the oyster-infecting parasite Perkinsus marinus. Parasitol. Int. 69: 13-16 (2018). PMID 30389616. [link to RRC of NBRP]

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

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QC test results

RIKEN BRC has sequenced portions of this material for quality test.
Please review the QC test results indicated by icon as well as clone information before placing your order.

Test sheet	RDB15790_A8E8p1.pdf

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

Primer: SV40pA3' element Reverse Sequence file: RDB15790_A8E8a.seq
>15790_15790_A8E8_2_SV40pA3'ElementReverse_F02_17_ABI24.ab1     1 TGGGGGGGGG GGCGGGAAAC CTATGGGGAA AAACGCCAGC AACGCGGGCC TTTTTACGGT    61 TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTTCTTTCC TGCGTTATCC CCTGATTCTG   121 TGGATAACCG TATTACCGCC TTTGAGTGAG CTGATACCGC TCGCCGCAGC CGAACGACCG   181 AGCGCAGCGA GTCAGTGAGC GAGGAAGCGG AAGAGCGCCC AATACGCAAA CCGCCTCTCC   241 CCGCGCGTTG GCCGATTCAT TAATGCAGCT GGCACGACAG GTTTCCCGAC TGGAAAGCGG   301 GCAGTGAGCG CAACGCAATT AATGTGAGTT AGCTCACTCA TTAGGCACCC CAGGCTTTAC   361 ACTTTATGCT TCCGGCTCGT ATGTTGTGTG GAATTGTGAG CGGATAACAA TTTCACACAG   421 GAAACAGCTA TGACCATGAT TACGCCAAGC TCAGAATTAA CCCTCACTAA AGGGACTAGT   481 CCTGCAGGTT TAAACGAATT CGCCCTTCTC GTAATGAGCC CAACCATTAT CGGATCGGTC   541 TTGCGGGCGA ACGAACATCT GCGGCAGGTG AGAGGTGGGA GTTTCCGGAT GCGGTCCGAG   601 TCATCGGCGA TGGAACTCCC CACAGGCCAG TGTGACGGTG AGCACTGATG TTCGATGGAG   661 ACCAAAGTGG CAGGGTTCGT ACTTCCCAGC CATGGTTGCG CCGTTTGTAA TTGATGTGAT   721 GGCAGAGCTG ACCGGTAGCA GTAGAAGGTG CGTTGATTCA GCTCAACAGC TCTCTTGTGT   781 TAGCAGGAAA AGTAGAAGCA GATCTAGGAT GATTCAGGAA ATCCCCTCAT CTTAGCGGTA   841 GTATGTTACT GTGCACCCAT CATGCCCATG GATGATCCGG GATCCCTTGT TGGGGTGAGG   901 TAGGTCCCTT TGGTGAGTTG GGATGATTGT GGGATCTGCC GATGGGCAGA AGATCAATGG   961 GCTCCATTCC CCCATCGGGG ACCCTCACGG GTAATCACTG AATCATTTTC AATGGGCTCA  1021 AAGGGGCGGG GCTTTGGTCG ACTTTCGACA AGTGGGGGTG GGGGACTGCG GTGAGCAAAG  1081 GGTGCCTTTG GCCACTTGGC CGAACCCAAT CGCTTTTGGC TTCCGGGCTT GTAATTGAAC  1141 CTAACGGGGA AGCCAGGAAA TTAGGGCCCG GGGGGCTATA AAATGGGGGA GACGGCAAAC 1201 TCACATCCAG //
Primer: M13_-40 Sequence file: RDB15790_A8E8b.seq
>15790_15790_A8E8_2_M13(-40)_G02_20_ABI24.ab1     1 ACCCAAGGGA AATGGTATAC GACTCACTAT AGGGCGAATT GAATTTAGCG GCCGCGAATT    61 CGCCCTTGGA GGACTTGAGG CTCTGTGACT ATACTCTTGA GATGAGGAAG ACTATGGTCT   121 TCTTACAGAA ACTCAGCCGT GGCATGATAG CGGAGTAAAA GCCTTCATGT TGGTTTCCGA   181 CCATGTTGTT ATCCATAGTC ACAACAAGCG TCGGAATAAC CATGGGATGC TCCTGGACGC   241 GCTGAGGGGG TTGAACCTTC TGATCAATCG TGGTAAGCTT AGGAGGATCC GCACTGTTTA   301 ATCATCTTCG CAGGGGCCAG TCTCCGAGTT GGAAGGGCCC AAGCAGCGGT GATCGCTGAA   361 TGCAAGAAGG CAATCAAGAA CAAGGATGCG GCTACCCTAG TTGGTGAGTC AAGTATGACT   421 GCCGTCGGCC ATTCCGGGTC TGCAGGTGTG ATCTCGCAGG GGGGCAATCT TGGTCCATCA   481 GCATGATGTT GATGTGCTTA TCTTTCATCA TCATTATCAT TGATTATGTG TGATAGTCTC   541 AATCATTAGA GTGCACTTCT TTGAGGGTAC TCGAGCCGAC CGAGTACACA CCTCTTTTCC   601 GTCAAAAAAT TACAATGGAT TGCGCCTGCT AACTATGACG GAGAGAAGAC TCAACTATAA   661 ACGTCTGTGG ATATCTACAT TCTGAGGACC AGTGGGCATC CGGGCAGGGT CAGTCCGGAG   721 GTATGTTAGG CAGGTCAGCC TCCTTGTAAA TCAATGAAGG ATTTGTGTTA TCGGCTGCTG   781 CTATTCCAAG CGCAGCTCAG TACGGAAATT ACGCGAGGAT GAAACAAAGT TGGGTTCATC   841 GAGCTGGTGA AATGAGAGTA ATGGTGTTTT CTGAAATGGG TAGATACACA GGTTGTTATA   901 TACAATTGGC CACTGGCACT CTCCCAAACG CAAACAAGAA CAAATGGAAG CGAGAACTTC   961 TCTGTAGTAT CGTTCACGGT TACTTGAATC CCTAACATGG CACAGATTCC CGAAGGAGAG  1021 AGGCCTTCTT ATACGTATTC TCCGACATGA TGACTCGTTA ACTCGGAGAT ACAAACCAAG 1081 G //
Primer: moe3_055R Sequence file: RDB15790_A8E8c.seq
>15790_15790_A8E8_2_moe3_055R_A03_03_ABI24.ab1     1 CAAGGGTTGA ATTCCGAACA TTGATGTAGA TCTCAACTAG ATGTGTTGTC TTAGGCACCG    61 GGCTTGCGGG TCATGCACCA GGTGCGCGGT CCTTCGGGCA CCTCGACGTC GGCGGTGACG   121 GTGAAGCCGA GCCGCTCGTA GAAGGGGAGG TTGCGGGGCG CGGAGGTCTC CAGGAAGGCG   181 GGCACCCCGG CGCGCTCGGC CGCCTCCACT CCGGGGAGCA CGACGGCGCT GCCCAGACCC   241 TTGCCCTGGT GGTCGGGCGA GACGCCGACG GTGGCCAGGA ACCACGCGGG CTCCTTGGGC   301 CGGTGCGGCG CCAGGAGGCC TTCCATCTGT TGCTGCGCGG CCAGCCGGGA ACCGCTCAAC   361 TCGGCCATGC GCGGGCCGAT CTCGGCGAAC ACCGCCCCCG CTTCGACGCT CTCCGGCGTG   421 GTCCAGACCG CCACCGCGGC GCCGTCGTCC GCGACCCACA CCTTGCCGAT GTCGAGCCCG   481 ACGCGCGTGA GGAAGAGTTC TTGCAGCTCG GTGACCCGCT CGATGTGGCG GTCCGGGTCG   541 ACGGTGTGGC GCGTGGCGGG GTAGTCGGCG AACGCGGCGG CGAGGGTGCG TACGGCCCGG   601 GGGACGTCGT CGCGGGTGGC GAGGCGCACC GTGGGCTTGT ACTCGGTCAT AGGACCGGGG   661 TTTTCTTCCA CGTCTCCTGC TTGCTTTAAC AGAGAGAAGT TCGTGGCTCC GCTTCCCTTG   721 TACAGCTCGT CCATGCCGAG AGTGATCCCG GCGGCGGTCA CGAACTCCAG CAGGACCATG   781 TGATCGCGCT TCTCGTTGGG GTCTTTGCTC AGCTTGGACT GGGTGCTCAG GTAGTGGTTG   841 TCGGGCAGCA GCACGGGGCC GTCGCCGATG GGGGTGTTCT GCTGGTAGTG GTCGGCGAGC   901 TGCACGCTGC CGTCCTCGAT GTTGTGGCGG ATCTTGAAGT TCACCTTGAT GCCGTTCTTC   961 TGCTTGTCGG CCATGATATA GACGTTGTGG GCTGTTGTAG TTGTACTCCA GCTTTGTGCC  1021 CCAGGATGTT TGCCGTCCTC CCTTGAAAGT CGATGCCCTT TCAGCTCGAT GCGATTCACA  1081 AGGGGTGTCG CCCCTCGACT CAACCTTCGG CGCGGATCTT GTAGTTGGCC GGTTCGATCC 1141 TGTGAG //
Primer: EGFP-C-rev Sequence file: RDB15790_A8E8d.seq
>15790_15790_A8E8_2_EGFP-C_rev_H02_23_ABI24.ab1     1 ACTCTGGATT GGGGTCTTTG CTCAGCTTGG ACTGGGTGCT CAGGTAGTGG TTGTCGGGCA    61 GCAGCACGGG GCCGTCGCCG ATGGGGGTGT TCTGCTGGTA GTGGTCGGCG AGCTGCACGC   121 TGCCGTCCTC GATGTTGTGG CGGATCTTGA AGTTCACCTT GATGCCGTTC TTCTGCTTGT   181 CGGCCATGAT ATAGACGTTG TGGCTGTTGT AGTTGTACTC CAGCTTGTGC CCCAGGATGT   241 TGCCGTCCTC CTTGAAGTCG ATGCCCTTCA GCTCGATGCG GTTCACCAGG GTGTCGCCCT   301 CGAACTTCAC CTCGGCGCGG GTCTTGTAGT TGCCGTCGTC CTTGAAGAAG ATGGTGCGCT   361 CCTGGACGTA GCCTTCGGGC ATGGCGGACT TGAAGAAGTC GTGCTGCTTC ATGTGGTCGG   421 GGTAGCGGCT GAAGCACTGC ACGCCGTAGG TCAGGGTGGT CACGAGGGTG GGCCAGGGCA   481 CGGGCAGCTT GCCGGTGGTG CAGATGAACT TCAGGGTCAG CTTGCCGTAG GTGGCATCGC   541 CCTCGCCCTC GCCGGACACG CTGAACTTGT GGCCGTTTAC GTCGCCGTCC AGCTCGACCA   601 GGATGGGCAC CACCCCGGTG AACAGCTCCT CGCCCTTGCT CACTCCGGAT CCGAGCTCCA   661 TATGCACTTT GTGATTCATG ATTGCGTTGA ACAGCGGGAT AGAGATTCAA GAGTGACGCG   721 AGTTCATCAA GAGTTCCACA CAACTCACTT AGCTATGTGT ATGCGTTCAA TGCGTCCGGT   781 CGATAGACAT CGTTATAGCG TCTCGCCTTA CAACATAGAG TCCATTTAGA AGGATTCAGC   841 GTTACGAAAT CAAGTGACTC GAGATTATGA TTTCCAAGCT AGTGCGAGAT TCGTGGAGAA   901 ATGCCCTTTG ATGGACAAGA CTGTGTTGCT CCGCGATACA TACTCACGCA TACAATGAAA   961 GGGAGCGTGA AGGGATATCT GTTGATGTCT ACTAGCAGAA CCCGTCCCTC CGCTCCTAAG  1021 TTCTCTAGCT CGCTGACGCC TCAAGGGTTC TAGTGAAGAT TTCGACTCGA GGATGCACAT  1081 CGTGGGAAGC ACTGCTTCGG AACGAGACCG AAATCCGAAC ATAAAAATTC TTAATGGAAT 1141 TTTGG //
Primer: Neo-M_R Sequence file: RDB15790_A8E8e.seq
>15790_15790_A8E8_2_Neo-M_R_B03_06_ABI24.ab1     1 CTCTCCAATT GCAGCCAGTC CCTTCCGCTT CAGTGACAAC GTCGAGCACA GCTGCGCAAG    61 GAACGCCCGT CGTGGCCAGC CACGATAGCC GCGCTGCCTC GTCTTGCAGT TCATTCAGGG   121 CACCGGACAG GTCGGTCTTG ACAAAAAGAA CCGGGCGCCC CTGCGCTGAC AGCCGGAACA   181 CGGCGGCATC AGAGCAGCCG ATTGTCTGTT GTGCCCAGTC ATAGCCGAAT AGCCTCTCCA   241 CCCAAGCGGC CGGAGAACCT GCGTGCAATC CATCTTGTTC AATCATGCGA AACGATCCTC   301 ATCCTGTCTC TTGATCAGAG CTTGATCCCC TGCGCCATCA GATCCTTGGC GGCAAGAAAG   361 CCATCCAGTT TACTTTGCAG GGCTTCCCAA CCTTACCAGA GGGCGCCCCA GCTGGCAATT   421 CCGGTTCGCT TGCTGTCCAT AAAACCGCCC AGTCTAGCTA TCGCCATGTA AGCCCACTGC   481 AAGCTACCTG CTTTCTCTTT GCGCTTGCGT TTTCCCTTGT CCAGATAGCC CAGTAGCTGA   541 CATTCATCCG GGGTCAGCAC CGTTTCTGCG GACTGGCTTT CTACGTGAAA AGGATCTAGG   601 TGAAGATCCT TTTTGATAAT CTCATGCCTG ACATTTATAT TCCCCAGAAC ATCAGGTTAA   661 TGGCGTTTTT GATGTCATTT TCGCGGTGGC TGAGATCAGC CACTTCTTCC CCGATAACGG   721 AGACCGGCAC ACTGGCCATA TCGGTGGTCA TCATGCGCCA GCTTTCATCC CCGATATGCA   781 CCACCGGGTA AAGTTCACGG GAGACTTTAT CTGACAGCAG ACGTGCACTG GCCAGGGGGA   841 TCACCATCCG TCGCCCCGGC GTGTCAATAA TATCACTCTG TACATCCACA AACAGACGAT   901 AACGGCTCTC TCTTTTATAG GTGTAAACCT TAAACTGCCG TACGTATAGG CTGCGCAACT   961 GTTGGGAAAG GGCGATCGGT GCGGGCCTCT TTCGCTATTA CGCCAGCTGG GCGAAGGGGG  1021 ATGTGCTGCA AAGGCGATTT AAGTTGGGTA ACGCCCAGGG GTTTTTCCCA GTCACGACGT  1081 TGTAAAACGA CGGCCAGTGG ATGTAATTAC GAACTCCACT ATAAGGGCGG AATTGGAATT 1141 TTAAGCCGGG TG //

Please visit Sequencing and PCR primers for primer information.

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References

Original, user report and related articles

original	Sakamoto, H., A Novel 2A-peptide-containing Plasmid to Generate Stable Perkinsus marinus Cells Expressing Organelle-targeted Genes. J. Eukaryot. Microbiol. 68 (5): e12861 (2021). PMID 34051022. [link to RRC of NBRP]
original	Sakamoto, H., Puromycin selection for stable transfectants of the oyster-infecting parasite Perkinsus marinus. Parasitol. Int. 69: 13-16 (2018). PMID 30389616. [link to RRC of NBRP]

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External Database
Perkinsus marinus MOE

Aequorea victoria GFP

Streptomyces alboniger puromycin-N-acetyl-transferase (pac)

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