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

CS-RfA-EG

Backbone vector plasmid of shRNA expressing lentivirus construct by Gateway(R) cloning with GFP marker driven by EF-1 alpha promoter.

Catalog number RDB04391
Resource name CS-RfA-EG
Clone info. Backbone vector plasmid of shRNA expressing lentivirus construct by Gateway(R) cloning with GFP marker driven by EF-1 alpha promoter.
Comment Commonly requested with pENTR4-H1 (RDB04395) for generation of shRNA expression vector and with pCMV-VSV-G-RSV-Rev (RDB04393) and pCAG-HIVgp (RDB04394) for packaging. Please note that Zeo resistance marker is not included in the lentivirus produced from this vector.
Vector backbone Plasmid
Selectable markers Please note that Zeo resistance marker is not included in the lentivirus produced from this vector. E. coli: Amp^r, Cm^r, ccdB.
Growth remarks Use DB3.1, ccdB Survival or equivalent to amplify this clone. 30 degC is fine.
Depositor|Developer Miyoshi, Hiroyuki |
Other clones in our bank

External Database

            Reference sequence
              
            Lentiviral plasmid list. [link]

            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 The availability of the BIOLOGICAL RESOURCE is limited to a RECIPIENT of a not-for profit institution for a not-for-profit academic purpose. The RECIPIENT agrees to expressly describe the late Dr. Hiroyuki Miyoshi as the Developer.
            Ordering Please visit Ordering instruction.[link] 
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            Material Transfer Agreement (MTA, for use for not-for-profit academic purpose)[Word]
            This RESOURCE contains a fluorescent protein owned or licensed by Global Life Sciences Solutions Operations UK Ltd. (Cytiva). Please refer to Schedule B and use Schedule A (download the Schedule A and Schedule B [link]).
            Remarks Remember that you will be working with samples containing infectious virus.
            Use Survival2 (Invitrogen) or equivalent to amplify this clone.
            提供案内 (日本国内) [open/close]

            提供条件 本件研究材料は、非営利機関の非営利学術研究に限って提供する。本件研究材料を利用した研究結果等を発表する際は、本件研究材料が故三好浩之博士により開発されたことを明示する。
            提供依頼 手続きの詳細は、「レンチウイルスベクターの提供申し込み」をご覧ください。
            提供依頼書 [Word]
            提供同意書 (非営利機関による非営利学術研究用)[Word]
            GFPリソースを含みます。Schedule Bをご一読の上Schedule A (英語版のみ) の提出をお願いします (Schedule AとSchedule Bを入手[link])。
            遺伝子組換え生物の受入れ確認書が必要です。当室にご請求ください。
            備考 このリソースはウイルス粒子産生用のため、取扱いに注意が必要です。
            このクローンの増殖にはSurvival2 (Invitrogen)あるいは同等の宿主を使用してください。

            Catalog # Resource name Shipping form Fee (non-profit org.)
            RDB04391 CS-RfA-EG DNA solution

            Ordering instruction of plasmids [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

            Sequence RDB04391z.seq
            Remarks, protocol and/or map (pdf) RDB04391.pdf

            Original reference

            original Miyoshi, H., Development of a self-inactivating lentivirus vector. J. Virol., 72 (10): 8150-8157 (1998). PMID 9733856.

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

            Featured content

            Featured content Lentivirus Vector (English text)
            Featured content Lentivirus Vector (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 RDB18667_B1Afp1-3.pdf check

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

            Primer: Cm_M_R
            Region: RfA(CmR,attR1),cTTP_CTS
            Sequence file: RDB18667_B1Afg.seq check
            >04391_18667_B1Af_1_Cm_M_R_B01_04_ABI24.ab1
                1 NNNNNNNNNN NNTCNNNNAT NNCCNGCTCN CCGTCTTTCN TTGCCNTACG GAATTCCGGA
               61 TGAGCATTCA TCAGGCGGGC AAGAATGTGA ATAAAGGCCG GATAAAACTT GTGCTTATTT
              121 TTCTTTACGG TCTTTAAAAA GGCCGTAATA TCCAGCTGAA CGGTCTGGTT ATAGGTACAT
              181 TGAGCAACTG ACTGAAATGC CTCAAAATGT TCTTTACGAT GCCATTGGGA TATATCAACG
              241 GTGGTATATC CAGTGATTTT TTTCTCCATT TTAGCTTCCT TAGCTCCTGA AAATCTCGAC
              301 GGATCCTAAC TCAAAATCCA CACATTATAC GAGCCGGAAG CATAAAGTGT AAAGCCTGGG
              361 GTGCCTAATG CGGCCGCCAT AGTGACTGGA TATGTTGTGT TTTACAGTAT TATGTAGTCT
              421 GTTTTTTATG CAAAATCTAA TTTAATATAT TGATATTTAT ATCATTTTAC GTTTCTCGTT
              481 CAGCTTTTTT GTACAAACTT GTGATAATTC AAAGTGAATT TCTGCTGTCC CTGTAATAAA
              541 CCCGAAAATT TTGAATTTTT GTAATTTGTT TTTGTAATTC TTTAGTTTGT ATGTCTGNTG
              601 CTATTATGTC TACTATTCNT TCCCCTGCAC TGTNNCCCCN NATCCCCCCT TTTCNTTTAA
              661 AATTGNGNAN GAANANNGCC NTTTGNANAT NCNNTCACNN ANCNAANNNN NCNNTCTCTG
            721 NNNNNCTNNN NNNNNNN
            //
            Primer: PRE_rev
            Region: PRE,EGFP 3'
            Sequence file: RDB18667_B1Afh.seq check
            >04391_18667_B1Af_1_PRE_rev_D01_10_ABI24.ab1
                1 NNNNNNNNNN NNNNNNNNGG ANCANCATAG TTAAGAATAC CAGTCAATCT TTCACAAATT
               61 TTGTAATCCA GAGGTTGATT ATCGATAAGC TTGATCCCTC GAGATCCTAC TAGTAGGATC
              121 TGAGTCCGGA CTTGTACAGC TCGTCCATGC CGAGAGTGAT CCCGGCGGCG GTCACGAACT
              181 CCAGCAGGAC CATGTGATCG CGCTTCTCGT TGGGGTCTTT GCTCAGGGCG GACTGGGTGC
              241 TCAGGTAGTG GTTGTCGGGC AGCAGCACGG GGCCGTCGCC GATGGGGGTG TTCTGCTGGT
              301 AGTGGTCGGC GAGCTGCACG CTGCCGTCCT CGATGTTGTG GCGGATCTTG AAGTTCACCT
              361 TGATGCCGTT CTTCTGCTTG TCGGCCATGA TATAGACGTT GTGGCTGTTG TAGTTGTACT
              421 CCAGCTTGTG CCCCAGGATG TTGCCGTCCT CCTTGAAGTC GATGCCCTTC AGCTCGATGC
              481 GGTTCACCAG GGTGTCGCCC TCGAACTTCA CCTCGGCGCG GGTCTTGTAG TTGCCGTCGT
              541 CCTTGAAGAA GANGGTGCGC TCCTGGACGT AGCCTTCGGG CATGGCGGAC TTGAAGAAGT
              601 CGTGCTGCTT CNTGTGGTCN GGGTAGCGGC TGAAGCACTG CNCGCCNTNN NCNNNGNTGG
              661 TCNCNANGGT GGGCCANGGN ACGGGCAGNT TGNNNGNGNN GNNNANGANN NTNNNGNNNN
            721 NNTTGNCNNN NNNNGNNNNN NNNCNNNNNC NNN
            //
            Primer: EF1a-N_R
            Region: EF-1 alpha pro,RfA(attR2,ccdB)
            Sequence file: RDB18667_B1Afi.seq check
            >04391_18667_B1Af_1_EF1a-N_R_A01_01_ABI24.ab1
                1 NNNNNNNNNN NNNNNNGGCG ANGTGCGCTC TGCCNNCTGA CGGGCACCGG ANCCTCACGA
               61 AGCTTGAATT ATCACCACTT TGTACAAGAA AGCTGAACGA GAAACGTAAA ATGATATAAA
              121 TATCAATATA TTAAATTAGA TTTTGCATAA AAAACAGACT ACATAATACT GTAAAACACA
              181 ACATATCCAG TCACTATGGT CGACCTGCAG ACTGGCTGTG TATAAGGGAG CCTGACATTT
              241 ATATTCCCCA GAACATCAGG TTAATGGCGT TTTTGATGTC ATTTTCGCGG TGGCTGAGAT
              301 CAGCCACTTC TTCCCCGATA ACGGAGACCG GCACACTGGC CATATCGGTG GTCATCATGC
              361 GCCAGCTTTC ATCCCCGATA TGCACCACCG GGTAAAGTTC ACGGGAGACT TTATCTGACA
              421 GCAGACGTGC ACTGGCCAGG GGGATCACCA TCCGTCGCCC GGGCGTGTCA ATAATATCAC
              481 TCTGTACATC CACAAACAGA CGATAACGGC TCTCTCTTTT ATAGGTGTAA ACCTTAAACT
              541 GCATTTCACC AGCCCCTGTT CTCGTCAGCA AAAGAGCCGT TCATTTCAAT AAACCGGGCG
              601 ACCTCAGCCA TCCCTTCCTG ANTTTCCGCT TTCCAGCGTT CGGCACGCAG ANGANGGGCT
              661 TCATTCTGCN TGGTTGNGCT TANNNNANCN GNANANANTG ACNTCATATA TGCNNNGAGC
            721 AACTGANANC NGTCNNNNNN
            //
            Primer: EF1a-C_F
            Region: EF-1 alpha pro,EGFP 5'
            Sequence file: RDB18667_B1Afj.seq check
            >04391_18667_B1Af_1_EF1a-C_F_C01_07_ABI24.ab1
                1 NNNNNNNNNN NNNTTTNANT TTGGNNCTTG GTTNNTTCTC NAGCCTCAGA CAGTGGTTCA
               61 AAGTTTTTTT CTTCCATTTC AGGTGTCGTG AACACGCTAC CGGTCGCCAC CATGGTGAGC
              121 AAGGGCGAGG AGCTGTTCAC CGGGGTGGTG CCCATCCTGG TCGAGCTGGA CGGCGACGTA
              181 AACGGCCACA AGTTCAGCGT GTCCGGCGAG GGCGAGGGCG ATGCCACCTA CGGCAAGCTG
              241 ACCCTGAAGT TCATCTGCAC CACCGGCAAG CTGCCCGTGC CCTGGCCCAC CCTCGTGACC
              301 ACCCTGACCT ACGGCGTGCA GTGCTTCAGC CGCTACCCCG ACCACATGAA GCAGCACGAC
              361 TTCTTCAAGT CCGCCATGCC CGAAGGCTAC GTCCAGGAGC GCACCATCTT CTTCAAGGAC
              421 GACGGCAACT ACAAGACCCG CGCCGAGGTG AAGTTCGAGG GCGACACCCT GGTGAACCGC
              481 ATCGAGCTGA AGGGCATCGA CTTCAAGGAG GACGGCAACA TCCTGGGGCA CAAGCTGGAG
              541 TACAACTACA ACAGCCACAA CGTCTATATC ATGGCCGACA AGCAGAAGAA CGGCATCAAG
              601 GTGAACTTCA AGATCCGCCA CAACATCGAG GANGGCAGCG TGCANCTCGC CNANCACTAN
              661 NNGCAGAANN NCCCNNTCGG NNANNNCCCC NNGCNGCTNN CCNANNNNNN NNACCNGANN
            721 NCCNNNNNNN NCNNNNNNNA NNNNCCNNNN NNNNNN
            //
            Primer: CMV_Forward
            Region: 5' LTR,HIV-1 psi
            Sequence file: RDB18667_B1Afk.seq check
            >04391_18667_B1Af_1_CMV_Forward_C01_03_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNTG NNNCNNNGGG CGGTAGGCGT GTACNGNGGG AGGTCTATAT
               61 AANCAGCGCG TNTTGCCTGT ACTGGGTCTC TCTGGTTAGA CCAGATCTGA GCCTGGGAGC
              121 TCTCTGGCTA ACTAGGGAAC CCACTGCTTA AGCCTCAATA AAGCTTGCCT TGAGTGCTTC
              181 AAGTAGTGTG TGCCCGTCTG TTGTGTGACT CTGGTAACTA GAGATCCCTC AGACCCTTTT
              241 AGTCAGTGTG GAAAATCTCT AGCAGTGGCG CCCGAACAGG GACTTGAAAG CGAAAGGGAA
              301 ACCAGAGGAG CTCTCTCGAC GCAGGACTCG GCTTGCTGAA GCGCGCACGG CAAGAGGCGA
              361 GGGGCGGCGA CTGGTGAGTA CGCCAAAAAT TTTGACTAGC GGAGGCTAGA AGGAGAGAGA
              421 TGGGTGCGAG AGCGTCAGTA TTAAGCGGGG GAGAATTAGA TCGCGATGGG AAAAAATTCG
              481 GTTAAGGCCA GGGGGAAAGA AAAAATATAA ATTAAAACAT ATAGTATGGG CAAGCAGGGA
              541 GCTAGAACGA TTCGCAGTTA ATCCTGGCCT GTTAGAAACA TCAGAAGGCT GTAGACAAAT
              601 ACTGGGACAG CTACAACCAT CCCTTCAGAC AGGATCAGAA GAACTTAGAT CATTATATAA
              661 TACAGTAGCA ACCCTCTATT GTGTGCATCA AAGGATAGAG ATAAAAGACA CCAAGGAAGC
              721 TTTAGACAAG ATAGAGGAAG AGCAAAACAA AAGTAAGANC ACCGCACAGC AAGCGGCCGC
              781 TGATCTTCAG ACCTGGAGGA NGAGATATGA NGGACAATTG GAGAAGTGAA TTATATAAAT
              841 ATAAAGTAGT AAAAATTGAA CCATTANGAG TAGCACCCAC CAAGGCAAAG ANAAGAGTGG
              901 TGCAGANAGA AAAAAGAGCA GNGGGAATAN GANCTTTGTT CCTNNGNNNC NNGGGNNCNN
            961 CNNNNANNCA CTATGGNNNC ANCGNNNNNG NNNCNGNNNN NNCNNNNCNN NNNNNNNN
            //
            Primer: HIVgp_R1
            Region: HIV-1 psi
            Sequence file: RDB18667_B1Afl.seq check
            >04391_18667_B1Af_1_HIVgp_R1_E01_13_ABI24.ab1
                1 NNNNNNNNNN NNNNNNCTCT TTTTTCTCTC TGCACCNCTC TTCTCTTTGC CTTGGTGGGT
               61 GCTACTCCTA ATGGTTCAAT TTTTACTACT TTATATTTAT ATAATTCACT TCTCCAATTG
              121 TCCCTCATAT CTCCTCCTCC AGGTCTGAAG ATCAGCGGCC GCTTGCTGTG CGGTGGTCTT
              181 ACTTTTGTTT TGCTCTTCCT CTATCTTGTC TAAAGCTTCC TTGGTGTCTT TTATCTCTAT
              241 CCTTTGATGC ACACAATAGA GGGTTGCTAC TGTATTATAT AATGATCTAA GTTCTTCTGA
              301 TCCTGTCTGA AGGGATGGTT GTAGCTGTCC CAGTATTTGT CTACAGCCTT CTGATGTTTC
              361 TAACAGGCCA GGATTAACTG CGAATCGTTC TAGCTCCCTG CTTGCCCATA CTATATGTTT
              421 TAATTTATAT TTTTTCTTTC CCCCTGGCCT TAACCGAATT TTTTCCCATC GCGATCTAAT
              481 TCTCCCCCGC TTAATACTGA CGCTCTCGCA CCCATCTCTC TCCTTCTAGC CTCCGCTAGT
              541 CAAAATTTTT GGCGTACTCA CCAGTCGCCG CCCCTCGCCT CTTGCCGTGC GCGCTTCAGC
              601 AAGCCGAGTC CTGCGTCGAG AGAGCTCCTC TGGTTTCCCT TTCNCTTTCN AGTCCCTGTT
              661 CNGGCGNNNC NGCTANAGAN TTTNCNCACT GANTAAANNN NCTGNNGNNN CTCTANNNAN
            721 NNNANTCNCN CNNNNNANNG NNNNNNNNNN
            //
            Primer: bGH_rev3
            Region: bGH pA,3' LTR
            Sequence file: RDB18667_B1Afm.seq check
            >04391_18667_B1Af_1_bGH_rev3_F01_16_ABI24.ab1
                1 NNNNNNNNNN NNNGGNNGNN GGGCNNACNA CAGATGGCTG GCAACTAGAA GGCACAGTCG
               61 AGGCTGATCA GCGGGTTTAA ACGGGCCCTG CTAGAGATTT TCCACACTGA CTAAAAGGGT
              121 CTGAGGGATC TCTAGTTACC AGAGTCACAC AACAGACGGG CACACACTAC TTGAAGCACT
              181 CAAGGCAAGC TTTATTGAGG CTTAAGCAGT GGGTTCCCTA GTTAGCCAGA GAGCTCCCAG
              241 GCTCAGATCT GGTCTAACCA GAGAGACCCA GTACAGTCCG GATGCAGCTC TCGGGCCATG
              301 TGATGAAATG CTAGGCGGCT GTCAAACCTC CACTCTAATA CTTCTCTCTC CGGGTCATCC
              361 ATCCCATGCA GGCTCACAGG GTGTAACAAG CGGGTGTTCT CTCCTTCATT GGCTTCTTCT
              421 ACCTTCTCTT GCTCAACTGG TACTAGCTTG TAGCACCATC CAAAGGTCAG TGGATATCTG
              481 ATCCCTGGCC CTGGTGTGTA GTTCTGCCAA TCAGGGAAGT AGCCTTGTGT GTGGTAGATC
              541 CACAGATCAA GGATATCTTG TCTTCGTTGG GAGTGAATTA GCCCTTCCNG TCCCCCCTTT
              601 TCTTTTAAAA AGTGGCTAAN ATCTACAGCT GCCTTGTAAG TCNTTGGTCT TAAAGGNANC
              661 TGANGTGTGA NTGNAAAANC NNNCTNCTCC TNCTCTNGNG CNTNTANNNN NGCNCNNNCN
            721 NNANTNGNNA NCTGCNNNNN NNNNN
            //
            Primer: SV40pro_ori_F
            Region: SV40 pro_ori,Zeo
            Sequence file: RDB18667_B1Afn.seq check
            >04391_18667_B1Af_1_SV40pro_ori_F_G01_19_ABI24.ab1
                1 NNNNNNGNNN NNGCCGCCTC TGCCTCTGAN CTATTCCNGA AGTAGTGAGG AGGCTTTTTT
               61 GGAGGCCTAG GCTTTTGCAA AAAGCTCCCG GGAGCTTGTA TATCCATTTT CGGATCTGAT
              121 CAGCACGTGT TGACAATTAA TCATCGGCAT AGTATATCGG CATAGTATAA TACGACAAGG
              181 TGAGGAACTA AACCATGGCC AAGTTGACCA GTGCCGTTCC GGTGCTCACC GCGCGCGACG
              241 TCGCCGGAGC GGTCGAGTTC TGGACCGACC GGCTCGGGTT CTCCCGGGAC TTCGTGGAGG
              301 ACGACTTCGC CGGTGTGGTC CGGGACGACG TGACCCTGTT CATCAGCGCG GTCCAGGACC
              361 AGGTGGTGCC GGACAACACC CTGGCCTGGG TGTGGGTGCG CGGCCTGGAC GAGCTGTACG
              421 CCGAGTGGTC GGAGGTCGTG TCCACGAACT TCCGGGACGC CTCCGGGCCG GCCATGACCG
              481 AGATCGGCGA GCAGCCGTGG GGGCGGGAGT TCGCCCTGCG CGACCCGGCC GGCAACTGCG
              541 TGCACTTCGT GGCCGAGGAG CAGGACTGAC ACGTGCTACG AGATTTCGAT TCCACCGCCG
              601 CCTTCTATGA AAGGNTGGGC TTCGGANTCG TTTTCCGGGA NGCCGGCTGG ANGATCCTCC
              661 NGCGCGGGGN NCTCNTGCTG GANTTCTTCG CCCACCCNNN NNNGNTTATN GNNNCTTNNN
            721 NNGGNNNNNA NTAANNCNNN NNNNNNN
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Miyoshi, H., Development of a self-inactivating lentivirus vector. J. Virol., 72 (10): 8150-8157 (1998). PMID 9733856.
            user_report Moriuchi, T., SUMOylation of RepoMan during late telophase regulates dephosphorylation of lamin A. J. Cell Sci. 134 (17): jcs247171 (2021). PMID 34387316.
            user_report Kawasaki, N., TUFT1 interacts with RABGAP1 and regulates mTORC1 signaling Cell Discovery 4 (1): (2018). PMID 29423269.
            user_report Yoshitomi, H., Human Sox4 facilitates the development of CXCL13-producing helper T cells in inflammatory environments. Nat. Commun. 9 (1): 3762 (2018). PMID 30232328.
            user_report Ishibashi, M., CD200-positive cancer associated fibroblasts augment the sensitivity of Epidermal Growth Factor Receptor mutation-positive lung adenocarcinomas to EGFR Tyrosine kinase inhibitors. Sci. Rep. 7: 46662 (2017). PMID 28429795.
            user_report Kobayashi, S., TGF-beta induces the differentiation of human CXCL13-producing CD4(+) T cells. Eur. J. Immunol. 46 (2): 360-371 (2016). PMID 26541894.
            user_report Katsura, A., MicroRNA-31 is a positive modulator of endothelial-mesenchymal transition and associated secretory phenotype induced by TGF-beta. Genes Cells 21 (1): 99-116 PMID 26663584.
            user_report Takahashi, Y., Perilipin2 plays a positive role in adipocytes during lipolysis by escaping proteasomal degradation. Sci Rep. 6: 20975 (2016). PMID 26876687.
            user_report Inoue, J., Identification of BCL11B as a regulator of adipogenesis. Sci. Rep. 6: 32750 (2016). PMID 27586877.
            user_report Neri, S., Podoplanin-expressing cancer-associated fibroblasts lead and enhance the local invasion of cancer cells in lung adenocarcinoma. Int. J. Cancer. 137 (4): 784-796 (2015). PMID 25648219.
            user_report Tanaka, Y., Dual Function of Histone H3 Lysine 36 Methyltransferase ASH1 in Regulation of Hox Gene Expression. PLoS One., 6 (11): e28171 (2011). PMID 22140534.
            user_report Ohoka, N., The orphan nuclear receptor RORalpha restrains adipocyte differentiation through a reduction of C/EBPbeta activity and perilipin gene expression. Mol. Endocrinol., 23, 759-771 (2009). PMID 19324970.
            user_report Naka, H., Requirement for COUP-TFI and II in the temporal specification of neural stem cells in CNS development. Nat. Neurosci. 11 (9): 1014-1023 (2008). PMID 19160499.
            user_report Fumoto, T., Orphan nuclear receptor Nur77 accelerates the initial phase of adipocyte differentiation in 3T3-L1 cells by promoting mitotic clonal expansion. J. Biochem. 141 (2): 181-192 (2007). PMID 17167038.
            user_report Tsukamoto H., TCR ligand avidity determines the mode of B-Raf/Raf-1/ERK activation leading to the activation of human CD4+ T cell clone. Eur. J. Immunol., 36, 1926-1937 (2006). PMID 16791876.

            2022.06.20

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