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pUC18K_ePAG2

Cloning vector harboring counterselection marker Escherichia coli phenylalanyl-tRNA synthetase a-subunit (ePheS).

Catalog number RDB17947
Resource name pUC18K_ePAG2
Clone info. Cloning vector harboring counterselection marker Escherichia coli phenylalanyl-tRNA synthetase a-subunit (ePheS) when 4-chloro-phenylalanine (4CP) is present. Kanamycin resistance. See Fig. S2 of Miyazaki, K. Biotechniques, 58 (2): 86-88, 2015 for constructs.
Comment Causing host cell death in the presence of 1 to 10% (w/v) of 4-chlorophenylalanine.
Vector backbone pUC18 (plasmid)
Selectable markers Ampicillin
Gene/insert name Escherichia coli pheS genomic DNA E. coli kanamycin resistance gene genomic DNA
Depositor|Developer Miyazaki, Kentaro |
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External Database
Escherichia coli pheS E. coli kanamycin resistance gene

        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 set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature(s) designated by the DEPOSITOR is requested. (Miyazaki K. et al. Biotechniques, 58 (2): 86-88, 2015)
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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 (Miyazaki K. et al. Biotechniques, 58 (2): 86-88, 2015)。
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        RDB17947 pUC18K_ePAG2 DNA solution

        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 (full) RDB17947zzk01.seq provided by the depositor

        Original reference

        original Miyazaki, K., Molecular engineering of a PheS counterselection marker for improved operating efficiency in Escherichia coli. Biotechniques 58 (2): 86-88 (2015). PMID 25652032.

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

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        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 RDB17947_B0Bcp1-1.pdf check

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

        Primer: KanR-C_F
        Region: insert mid (KmR 3', pheS 5')
        Sequence file: RDB17947_B0Bca.seq check
        >D06895A5_B0Bc_1_kanR-C_F_B03_06_ABI24.ab1
            1 NNNNNNNNNN NNNNNNNNNN NNNNNNNGTA NTGNNAATCC TGATATGAAT AAATTGCAGT
           61 TTCATTTGAT GCTCGATGAG TTTTTCTAAA GGAGGTAAAA CACATGTCAC ATCTCGCAGA
          121 ACTGGTTGCC AGTGCGAAGG CGGCCATTAG CCAGGCGTCA GATGTTGCCG CGTTAGATAA
          181 TGTGCGCGTC GAATATTTGG GTAAAAAAGG GCACTTAACC CTTCAGATGA CGACCCTGCG
          241 TGAGCTGCCG CCAGAAGAGC GTCCGGCAGC TGGTGCGGTT ATCAACGAAG CGAAAGAGCA
          301 GGTTCAGCAG GCGCTGAATG CGCGTAAAGC GGAACTGGAA AGCGCTGCAC TGAATGCGCG
          361 TCTGGCGGCG GAAACGATTG ATGTCTCTCT GCCAGGTCGT CGCATTGAAA ACGGCGGTCT
          421 GCATCCGGTT ACCCGTACCA TCGACCGTAT CGAAAGTTTC TTCGGTGAGC TTGGCTTTAC
          481 CGTGGCAACC GGGCCGGAAA TCGAAGACGA TTATCATAAC TTCGATGCTC TGAACATTCC
          541 TGGTCACCAC CCGGCGCGCG CTGACCACGA CACTTTCTGG TTTGACACTA CCCGCCTGCT
          601 GCGTACCCAG ACCTCTGGCG TACAGATCCG CACCATGAAA GCCCAGCAGC CACCGATTCG
          661 TATCATCGCG CCTGGCCGTG TTTATCGTAA CGACTACGAC CAGACTCACA CGCCGATGTT
          721 CCATCAGATG GAAGGTCTGA TTGTTGATAC CAACATCAGC TTTACCAACC TGAAAGGCAC
          781 GCTGCACGAC TTCCTGCGTA ACTTCTTTGA GGAAGATTTG CAGANTCGCT TCCGTCCTTC
          841 CTACTTCCCG TTTGCTGANN CTTCTGCANA ANNGGACGTC NTGGGTAAAA ACNGTAAATG
          901 GCTGGAAGTG CTGGGCTGCG GGATNNNGNA TNCNANNGTG NTGCGTANCG TTGNNATCNA
          961 NCNNNAAGTT TACTCNGGNN TTNNGGNTCG GGATNGGGAT GGNNCGTCNN NNTATGNNGC
        1021 GNNNNNNNNN N
        //
        Primer: KanR-N_R
        Region: insert 5' (KmR 5', AmpR pro, MCS)
        Sequence file: RDB17947_B0Bcb.seq check
        >D06895A5_B0Bc_1_kanR-N_R_A03_03_ABI24.ab1
            1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNCATCN NNGTTGGAAT TTAATCGCGG
           61 CCTCGAGCAA GACGTTTCCC GTTGAATATG GCTCATACTC TTCCTTTTTC AATATTATTG
          121 AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA
          181 TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG TCTAAGAAAC
          241 CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC ACGAGGCCCT TTCGTCTCGC
          301 GCGTTTCGGT GATGACGGTG AAAACCTCTG ACACATGCAG CTCCCGGAGA CGGTCACAGC
          361 TTGTCTGTAA GCGGATGCCG GGAGCAGACA AGCCCGTCAG GGCGCGTCAG CGGGTGTTGG
          421 CGGGTGTCGG GGCTGGCTTA ACTATGCGGC ATCAGAGCAG ATTGTACTGA GAGTGCACCA
          481 TGTGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA TACCGCATCA GGCGCCATTC
          541 GCCATTCAGG CTGCGCAACT GTTGGGAAGG GCGATCGGTG CGGGCCTCTT CGCTATTACG
          601 CCAGCTGGCG AAAGGGGGAT GTGCTGCAAG GCGATTAAGT TGGGTAACGC CAGGGTTTTC
          661 CCAGTCACGA CGTTGTAAAA CGACGGCCAG TGCCAAGCTT GCATGCCTGC AGGTCGACTC
          721 TAGAGGATCC CCGGGTACCG AGCTCGANTT CGTAATCATG GTCATATGTG TTTCCTGTGT
          781 GAAATTGTTA TCCGCTCNCN ATTCCACACN ACATACNAGC CGGAANCATA AAGTGTAAAG
          841 CCTGGGGGNG CCTAATGAGT GAGCTNNCTC ACATTNNTTG CGTNGCGCNC NCTGCCNGCT
          901 TTCCANNNNG GNAANCNGTC NNGNCNNCTG CNTTAATGNA NCGGNNNACG CNNGGGGNNA
          961 NGNNGNTTGN GNANNNNGCG CTCTTCCGNT NNNNCGCNCN NNNANNNNNT NCNNNNNNNC
        1021 NTNNNNNNNC NNCNNNNNGN NANNNNNNCN NNNNANNNNN NNNNN
        //
        Primer: ePAG_R1
        Region: insert 3' (pheS 3')
        Sequence file: RDB17947_B0Bcc.seq check
        >D06895A5_B0Bc_1_ePAG_R1_C03_09_ABI24.ab1
            1 NNNNNNNNNN NNNNNNNNNN NNTNNCCTNN NNNNTTTTNN TTANNNNNGA AGTTTTAAAT
           61 CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGGAA TTATTATTTA AACTGTTTGA
          121 GGAAACGCAG ATCGTTTTCG AAGAATGAAC GCAGGTCGGT GACGCCGTAA CGCAACATAG
          181 TCAGACGCTC CATCCCCATC CCGAACCCGA AACCAGAGTA AACTTCCGGG TCGATGCCAA
          241 CGTTACGCAA CACGTTCGGA TGCACCATCC CGCAGCCCAG CACTTCCAGC CATTTACCGT
          301 TTTTACCCAT GACGTCCACT TCTGCAGAAG GTTCAGCAAA CGGGAAGTAG GAAGGACGGA
          361 AGCGAATCTG CAAATCTTCC TCAAAGAAGT TACGCAGGAA GTCGTGCAGC GTGCCTTTCA
          421 GGTTGGTAAA GCTGATGTTG GTATCAACAA TCAGACCTTC CATCTGATGG AACATCGGCG
          481 TGTGAGTCTG GTCGTAGTCG TTACGATAAA CACGGCCAGG CGCGATGATA CGAATCGGTG
          541 GCTGCTGGGC TTTCATGGTG CGGATCTGTA CGCCAGAGGT CTGGGTACGC AGCAGGCGGG
          601 TAGTGTCAAA CCAGAAAGTG TCGTGGTCAG CGCGCGCCGG GTGGTGACCA GGAATGTTCA
          661 GAGCATCGAA GTTATGATAA TCGTCTTCGA TTTCCGGCCC GGTTGCCACG GTAAAGCCAA
          721 GCTCACCGAA GAAACTTTCG ATACGGTCGA TGGTACGGGT AACCGGATGC AGACCGCCGT
          781 TTTCNATGCG ACGANCTGGC AGAGAGACAT CNANNCGTTT CCNCCGCCAG ANGCGCATTC
          841 AGTGCAGCGC TTTCCAGTTC CGCTTTACNC GCATTCAGCG NCNGCTGANN CTGCTCTTTC
          901 NCTNCNTTGA TAACCGCACN NGCTGCNNNA NGCTCNNCTN NNNNNAGCTC ACGCANGNNN
          961 GTCNTCTGNN NGGNNNANTG NCCTTTTTTA NNNNNNNNTN NNNNNGNNNA TNNTCTNNNG
        1021 NNGCANCNTC NNNNNNNNNN
        //
        Primer: SV40pA3'Element Reverse
        Region: ColE1 ori, MCS
        Sequence file: RDB17947_B0Bcd.seq check
        >17947_17947_B0Bc_1_SV40pA3'elementReverse_G01_07_ABI08.ab1
            1 NNNNNNNNNN NGNNNNNNNN NNNNNNNNNA NGCNNNNNNN NCGGCCTTTT TACGGTTCCT
           61 GGCCTTTTGC TGGCCTTTTG CTCACATGTT CTTTCCTGCG TTATCCCCTG ATTCTGTGGA
          121 TAACCGTATT ACCGCCTTTG AGTGAGCTGA TACCGCTCGC CGCAGCCGAA CGACCGAGCG
          181 CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA GCGCCCAATA CGCAAACCGC CTCTCCCCGC
          241 GCGTTGGCCG ATTCATTAAT GCAGCTGGCA CGACAGGTTT CCCGACTGGA AAGCGGGCAG
          301 TGAGCGCAAC GCAATTAATG TGAGTTAGCT CACTCATTAG GCACCCCAGG CTTTACACTT
          361 TATGCTTCCG GCTCGTATGT TGTGTGGAAT TGTGAGCGGA TAACAATTTC ACACAGGAAA
          421 CACATATGAC CATGATTACG AATTCGAGCT CGGTACCCGG GGATCCTCTA GAGTCGACCT
          481 GCAGGCATGC AAGCTTGGCA CTGGCCGTCG TTTTACAACG TCGTGACTGG GAAAACCCTG
          541 GCGTTACCCA ACTTAATCGC CTTGCAGCAC ATCCCCCTTT CGCCAGCTGG CGTAATAGCG
          601 AAGANGCCCG CACCGATCGC CCTTCCCAAC AGTTGCGCAN CCTGNATGGC NANNGGNGCC
          661 TGATGCGGNA TTTTCTCNTT ANNCATCTGN GCNNNATTTC ACANCGCANN TGGNGCNNTC
        721 TCNNNNCAAT CTGCTCNNNN NNNNNN
        //

        Please visit Sequencing and PCR primers for primer information.


        References

        Original, user report and related articles

        original Miyazaki, K., Molecular engineering of a PheS counterselection marker for improved operating efficiency in Escherichia coli. Biotechniques 58 (2): 86-88 (2015). PMID 25652032.

        2022.05.17

        GNP_filter3_RDBDEP_html_220516.pl