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pcDNA4c/His-Xpress-HA-hBIG2(WT)

Expression vector of human BIG2, wild type.

Catalog number RDB15364
Resource name pcDNA4c/His-Xpress-HA-hBIG2(WT)
Alternative name N7
Clone info. Expression vector of human Arf guanine nucleotide-exchange factors 2 (BIG2), wild type, tagged with Xpress and HA epitope at N-ter. This clone contains a cDNA fragment corresponding to the 180nt to 5972nt of NM_006420.3. Known differences (at least) : C to T substitution at 5342nt (Leu, synonym) ; A to G substitution at 5345nt (Lys, synonym).
Comment 1. BIG1 and BIG2 containing plasmids seem to be toxic to bacteria. After transformation, you need longer incubation time to get colonies as compared with usual transformed bacteria. Moreover, the colonies are smaller than usual ones. So, please take small colonies for liquid culture (The depositor is usually using DH5alpha for transformation).
2. Mini-prep: For liquid culture, you should use very fresh colonies otherwise you will get very low amounts of plasmids.
3. Maxi-prep (Large scale prep): Do not make a preculture from a single colony for large scale culture. You will get very low amounts of plasmids. The depositor would recommend you that after transformation and recovery culture, bring all transformed bacteria directly to large scale culture (200 ml or more of ampicillin containing medium). This helps a lot to get plasmids.
Expression was confirmed by the depositor with immunostaining.
Vector backbone pcDNA4/HisMax (plasmid)
Size of vector backbone 5.3 kb
Selectable markers Amp^r (E. coli), Zeo^r (mammalian cells).
Gene/insert name human BIG2 cDNA
Depositor|Developer Nakayama, Kazuhisa |
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External Database
human BIG2

          Reference sequence
            
           

          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 for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested (Ishizaki, R. et al. Mol. Biol. Cell, 19 (6): 2650-2660, 2008). RECIPIENT agrees to use the BIOLOGICAL RESOURCE only for academic research in the non-profit organization.
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          提供条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する (Ishizaki, R. et al. Mol. Biol. Cell, 19 (6): 2650-2660, 2008)。本件リソースは非営利機関にのみ提供し、利用範囲は学術研究に制限する。
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          Catalog # Resource name Shipping form Fee (non-profit org.)
          RDB15364 pcDNA4c/His-Xpress-HA-hBIG2(WT) DNA solution

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

          How to cite this biological resource

          Materials & Methods section:

          The pcDNA4c/His-Xpress-HA-hBIG2(WT) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15364).

          Reference section:

          Ishizaki, R., Shin, H.W., Mitsuhashi, H., Nakayama, K., Redundant roles of BIG2 and BIG1, guanine-nucleotide exchange factors for ADP-ribosylation factors in membrane traffic between the trans-Golgi network and endosomes. Mol. Biol. Cell 19 (6): 2650-2660 (2008). PMID 18417613.

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


          References and tips

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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 check icon as well as clone information before placing your order.

          Test sheet RDB15364_A7I5p1-1.pdf check

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

          Primer: CMV_Forward (Pr0016)
          Region: CMV pro, QBI SP163, 6xHis, T7 tag, Xpress tag, HA, insert 5'
          Sequence file: RDB15364_A7I5a.seq check
          >D03588D1_2_A7I5_CMV-Forward_C06_09_ABI24.ab1
              1 NNNNNNNNNN NNNTTGNNNC NNNGGGCGGT AGGCGTGTAC GGTGGGAGGT CTATATAAGC
             61 AGAGCTCTCT GGCTAACTAG AGAACCCACT GCTTACTGGC TTATCGAAAT TAATACGACT
            121 CACTATAGGG AGACCCAAGC TGGCTAGCGT TTAAACTTAA GCTTAGCGCA GAGGCTTGGG
            181 GCAGCCGAGC GGCAGCCAGG CCCCGGCCCG GGCCTCGGTT CCAGAAGGGA GAGGAGCCCG
            241 CCAAGGCGCG CAAGAGAGCG GGCTGCCTCG CAGTCCGAGC CGGAGAGGGA GCGCGAGCCG
            301 CGCCGGCCCC GGACGGCCTC CGAAACCATG GGGGGTTCTC ATCATCATCA TCATCATGGT
            361 ATGGCTAGCA TGACTGGTGG ACAGCAAATG GGTCGGGATC TGTACGACGA TGACGATAAG
            421 GTACCTACCA TGTACCCGTA CGACGTGCCC GACTACGCGG GATCCACCAT GCAGGANAGC
            481 CAGACCAAGA GCATGTTCGT GTCCCGGGCC CTGGANAAGA TCCTAGCCGA CAAGGANGTG
            541 AAGCGGCCCC AGCACTCCCA GCTGCGCANG GCCTGCCAGG NGGCGCTCNA TGAAATTAAA
            601 GCANAAATAG AAAANCANNN NNTNNGCACT GCTGCANNNN CNAANGNNAA ACTTCNTTGA
          661 ANCTGANNNG NNTTTTCNNN NNTNNNANCT ANCNNGNNNN TNNNNNNNCN NNNNNNNNN
          //
          Primer: BGH_rev2 (Pr0606)
          Region: BGH pA, insert 3'
          Sequence file: RDB15364_A7I5b.seq check
          >D03588D1_A7I5_2_BGH_rev2_D05_04_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NGGGGGNNGG GCANNNACNG ATGGCTGGCA ACTAGAAGGC
             61 ACAGTCGAGG CTGATCAGCG GGTTTAAACG GGCCCTCTAG ACTCGAGCGG CCGCAAACGT
            121 TTTTTTTTTT TTTTTTTATT TTAGGTGGCA GTTAGCCGTT ACAAGAAGTT TTATTAACAT
            181 ATTCTCTAAT AATCACAAAC CTATACAGCT ATGTTCAGTA TTTTCAAGGA GAGTGACTTC
            241 TTACAAAACG TTTATAGCCA ATCATGGGAT TTTGCTAGAA AACGATGGCA TATGCAATGA
            301 CAGATTTGAC AGTTTAATCA NAAAAATGGA ATGATAGAAA TGAAGCANAC TCCCCACCTA
            361 CTGTGCTAAC TGGGTATGCA ACAGTTTAAT GAAATACCAT AAGCCTTAGT TGGAGACTGT
            421 GATGGAACAG CGTANAGGCC ATCCGTTTCC AGGCCATTCT GAGATCCAAG ATGCCAACTC
            481 CTTGTTTCTA TGAAACTTCA CGAGATGTGC CAGTCAGAAA TGGCATGCTG AACATTCTGC
            541 AGAGCTGCAG CACTGGCCTG GGCAGCCAGG GCTACCACAC TGGTGACAGT GCTGCTGGTA
            601 CCTGTGATGG CTCTTCTGGT ATCCATATCT TATACACAAC ACCTATCCGT AGGAAGAACT
            661 TCCGCANAAC TGCTCGGAGC TCANGGATCA GGTCAAACTG CATAATTTCA CACAAGTAGG
            721 GGTAGTACAT TGAAGCATGT GCTTTGAACT TTTCATCATT TATCTTANNG GTTTTAGTTA
            781 GAAGTAACNA CAAGAGANTT GTCCAGGCCT NCCGATGGCT CTCANAATTC ACAGNGNTGA
            841 AATAGGNNAN ANCTTCNNTG CAAACNGTTN AAAGTCTCTG CTGNATTTCT NNCNNNGANN
            901 CCNNGCNGNT CNNATNNNNA TACATTNNAA ANNGGNNNNT CAAANNNNNN NCNNNGNNNN
          961 NNNNTNNNNN NTNNNNNNNN NGGNNNNNNN NTNNNNNNNN NN
          //
          Primer: hBIG2_F1 (Pr0344)
          Region: insert mid (E738 WT)
          Sequence file: RDB15364_A7I5c.seq check
          >D03588D1_A7I5_2_hBIG2_F1_C02_03_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNT TCTCNNGGNN CAGGGCATGC TGGGAACGTC AGTTGAAGAC
             61 ATAGCCCAAT TCCTGCACCA GGAGGAGCGC CTGGATTCCA CCCAAGTAGG CGATTTTCTG
            121 GGAGATAGCG CAAGGTTCAA CAAGGAGGTG ATGTATGCCT ACGTGGACCA ACTTGACTTC
            181 TGTGAAAAAG AATTTGTCTC AGCCCTGCGG ACATTCCTAG AAGGTTTCCG CCTACCTGGA
            241 GAAGCCCAAA AGATTGACCG ATTAATGGAG AAGTTTGCCG CAAGATACAT AGAATGCAAC
            301 CAAGGGCAAA CTCTGTTTGC TAGTGCTGAC ACTGCTTATG TCCTAGCGTA TTCAATTATT
            361 ATGCTGACTA CAGACTTGCA CAGTCCTCAG GTAAAAAATA AAATGACGAA AGAGCAGTAT
            421 ATTAAAATGA ATCGGGGTAT CAATGATAGT AAAGATCTGC CAGAAGAGTA TCTCTCAAGC
            481 ATCTATGAAG AGATAGAAGG CAAGAAAATT GCAATGAAAG AAACAAAAGA GCTAACGATT
            541 GCAACCAAAT CTACTAAGCA GAATGTAGCT AGTGAAAAGC AGCGGCGGCT GCTGTACAAC
            601 TTAGAGATGG AGCAAATGGC TAAAACAGCC AAAGCTCTGA TGGAGGCTGT GAGCCATGCC
            661 AAAGCCCCGT TTACCAGTGC CACTCACCTG GACCATGTCC GGCCAATGTT CNAACTGGTG
            721 TGGACGCCAC TATTGGCAGC CTACAGCATC GGACTCCAGA ACTGTGATGA CACTGAAGTG
            781 GCCTCCTTGT GTTTGGAANG CATCCGATGT GCAATCCGAN TCGCCTGCAT CTTTGGAATG
            841 CAGCTGGAAC GANATGCCTA TGTTCANGCT CTTGCTCGCT TCTCCNNACT CNNNGCNNGC
            901 TCNNGCATCN CAGAANTGNA NNANAAAAAC ATCNACNCCN TTAANANGCT TANCACANNG
          961 GCNCANNCCG ANGGNNNNNA NCNNNNGNNN NNNNNNN
          //
          Primer: hBIG2_R1 (Pr1019)
          Region: insert 5', HA, Xpress tag, T7 tag, 6xHis, QBI SP163
          Sequence file: RDB15364_A7I5d.seq check
          >D03588D1_A7I5_2_hBIG2_R1_E02_05_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNGNA GCCTNGGCAG TGGTTTGNTT GATGAGANTT TTGCTGGCCA
             61 AATAGATATT GTAACATGTT CTCACTGTCT GCAGGATAGT ACCCTCATGA ATTTCAATGT
            121 GTGGGGAAGT CACTGCAGTC AGAAGAGCCT TAATTATTTG TAACTGAACC CCTTCATCAG
            181 TCTGAGGGCC CTGAAAACAA CTGCAAATGG TTTCAACAAT TCTGTCGATC AGCCGCTTCC
            241 CAGGGGCTCC ACTGTCAGGG GCGTTGCCAG TGATGTGCCC GTATGCGATG AGTTTCTGCA
            301 AGCAGTCAAG GGATGTGCTG ACTACCCTTG GGGACTTGGA CTGGCAAGCT AGCTCGAATG
            361 GAAGAAAATA CTTGTCAGCT TCAATGAAGT TTGCCTTTGG TGGTGCAGCA GTGCCAAGCC
            421 TCTGCTTTTC TATTTCTGCT TTAATTTCAT CGAGCGCCAC CTGGCAGGCC CTGCGCAGCT
            481 GGGAGTGCTG GGGCCGCTTC ACCTCCTTGT CGGCTAGGAT CTTCTCCAGG GCCCGGGACA
            541 CGAACATGCT CTTGGTCTGG CTCTCCTGCA TGGTGGATCC CGCGTAGTCG GGCACGTCGT
            601 ACGGGTACAT GGTAGGTACC TTATCGTCAT CGTCGTACAG ATCCCGACCC ATTTGCTGTC
            661 CACCAGTCAT GCTAGCCATA CCATGATGAT GATGATGATG AGAACCCCCC ATGGTTTCGG
            721 ANGCCGTCCG GGGCCGGCGC GGCTCGCGCT CCCTCTCCGG CTCGGACTGC GANGCAGCCC
            781 GCTCTCTTGC GCGCCTTGNC GGGCTCCTCT CCCTTCTGGA ACCGAGGCCC GGGCCGGGGC
            841 CTGNCTGCCG CTCGGCTGCC CNNAGCCTCT GCGCTAANCT TAAGTTTAAA CGCTAGCCAG
            901 CTNNNNNNTN CCNATAGTGA GTNGNANTAA TTTCNATNAG CCAGTNANCA GNGGGNNCTC
          961 NANTNANNNN NANNNCTCNG NNNANNNNNN NNNN
          //
          Primer: hBIG2_F2 (Pr1020)
          Region: insert 3'
          Sequence file: RDB15364_A7I5e.seq check
          >D03588D1_A7I5_2_hBIG2_F2_F02_06_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNTCTTC NNNNNNAGAA ACCAGCAGCC TGGCCTGTTG
             61 TTTGAGGNNC CTGTTTCGAA TGTATGTTGA TGAGAACCGC AGGGNTTCCT GGGAAGAAAT
            121 ACAGCAGAGA CTTTTAACTG TTTGCAGTGA AGCTCTTGCC TATTTCATCA CTGTGAATTC
            181 TGAGAGCCAT CGGGAGGCCT GGACAAGTCT CTTGTTGTTA CTTCTAACTA AAACCCTTAA
            241 GATAAATGAT GAAAAGTTCA AAGCACATGC TTCAATGTAC TACCCCTACT TGTGTGAAAT
            301 TATGCAGTTT GACCTGATCC CTGAGCTCCG AGCAGTTCTG CGGAAGTTCT TCCTACGGAT
            361 AGGTGTTGTG TATAAGATAT GGATACCAGA AGAGCCATCA CAGGTACCAG CAGCACTGTC
            421 ACCAGTGTGG TAGCCCTGGC TGCCCAGGCC AGTGCTGCAG CTCTGCAGAA TGTTCAGCAT
            481 GCCATTTCTG ACTGGCACAT CTCGTGAAGT TTCATAGAAA CAAGGAGTTG GCATCTTGGA
            541 TCTCAGAATG GCCTGGAAAC GGATGGCCTC TACGCTGTTC CATCACAGTC TCCAACTAAG
            601 GCTTATGGTA TTTCATTAAA CTGTTGCATA CCCAGTTAGC ACAGTAGGTG GGGAGTCTGC
            661 TTCATTTCTA TCATTCCATT TTTCTGATTA AACTGTCAAA TCTGTCATTG CATATGCCAT
            721 CGTTTTCTAG CAAAATCCCA TGATTGGCTA TAAACGTTTT GTAAGAAGTC ACTCTCCTTG
            781 AAAATACTGA ACATAGCTGT ATAGGTTTGT GANTATTAGA GAATATGTTA ATAAAACTTC
            841 TTGTAACGGC TANNTGCNAC CNNANNNAAA AAAAAAAAAA AAAAACGTTT GCGGNNGCTC
            901 GNNTCTANAG GNCCCGTTNN NNCCGCTNNN CANCNNCNNN NNNGNCTTNN ANTTGNNNNC
          961 NNTCNNNNNN TNNNCNNNNN NNNNN
          //
          Primer: SV40pro_F (Pr0110)
          Region: EM7 pro, ZeoR, SV40 pA
          Sequence file: RDB15364_A7I5f.seq check
          >D03588D1_A7I5_2_SV40pro_F_D02_04_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NGANNNGTGN NGNNGCTTTT TTGGANGCCT AGGCTTTTGC
             61 AAAAAGCTCC CGGGAGCTTG TATATCCATT TTCGGATCTG ATCAGCACGT GTTGACAATT
            121 AATCATCGGC ATAGTATATC GGCATAGTAT AATACGACAA GGTGAGGAAC TAAACCATGG
            181 CCAAGTTGAC CAGTGCCGTT CCGGTGCTCA CCGCGCGCGA CGTCGCCGGA GCGGTCGAGT
            241 TCTGGACCGA CCGGCTCGGG TTCTCCCGGG ACTTCGTGGA GGACGACTTC GCCGGTGTGG
            301 TCCGGGACGA CGTGACCCTG TTCATCAGCG CGGTCCAGGA CCAGGTGGTG CCGGACAACA
            361 CCCTGGCCTG GGTGTGGGTG CGCGGCCTGG ACGAGCTGTA CGCCGAGTGG TCGGAGGTCG
            421 TGTCCACGAA CTTCCGGGAC GCCTCCGGGC CGGCCATGAC CGAGATCGGC GAGCAGCCGT
            481 GGGGGCGGGA GTTCGCCCTG CGCGACCCGG CCGGCAACTG CGTGCACTTC GTGGCCGAGG
            541 AGCAGGACTG ACACGTGCTA CGAGATTTCG ATTCCACCGC CGCCTTCTAT GAAAGGTTGG
            601 GCTTCGGAAT CGTTTTCCGG GACGCCGGCT GGATGATCCT CCAGCGCGGG GATCTCATGC
            661 TGGAGTTCTT CGCCCACCCC AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA
            721 ATAGCATCAC AAATTTCACA AATAAAGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT
            781 CNNAACTCAT CAATGTATCT TATCATGTCT GTATACCGTC GACCTCTANC TAGAGCTTGG
            841 CGTANTCATG GTCATAGCTG TTTCCTGTGT GAAATTGTTA TCCGCTCANN NTTCCNCACA
            901 ACATACNANC NNNAAGCNTA AAGNGTAAAG CCNGGGGGNN NCNNATGAGT GANCTAACTC
          961 ACNTTNANNG CGNNGNNNNN NCNNNCNNCT TTCCANNNNG NNANNNNNNN NNN
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

          original Ishizaki, R., Redundant roles of BIG2 and BIG1, guanine-nucleotide exchange factors for ADP-ribosylation factors in membrane traffic between the trans-Golgi network and endosomes. Mol. Biol. Cell 19 (6): 2650-2660 (2008). PMID 18417613.

          2023.05.01

          GNP_filter3_RDBDEP_html_230501.pl