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

pUC18K_mPAG2

Cloning vector harboring PheS counterselection marker. Kanamycin resistance.

Catalog number RDB18372
Resource name pUC18K_mPAG2
Clone info. Cloning vector harboring counterselection codon-modified marker Escherichia coli phenylalanyl-tRNA synthetase variant (T251A/A294G; mPAG2), a-subunit (ePheS) when 4-chloro-phenylalanine (4CP) is present. Kanamycin resistance. An improved clone of pUC18K_ePAG2 (RDB17947). The risk of recombination is lower.
Comment Version with reduce recombination against PheS gene in E. coli genome. A tendency to host cell death in the presence of 1-10% (w/v) 4-chlorophenylalanine.
Vector backbone pUC18 (plasmid)
Selectable markers Kanamycin, pheS
Gene/insert name Escherichia coli pheS genomic DNA
Depositor|Developer Suzuki, Kaoru | Miyazaki, Kentaro |
Other clones in our bank

External Database
Escherichia coli pheS

          Reference sequence
            
           
          Sequence (full) RDB18372zzk01.seq provided by the depositor

          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 literature designated by the DEPOSITOR is requested. (Miyazaki K. Biotechniques, 58 (2): 86-88, 2015)
          Ordering Please visit Information of Request for Distribution.[link] For for-profit-research purpose, please contact us. 
          Order form 
          [Credit Card Payment]  [Bank Transfer Payment]
          Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
          提供案内 (日本国内) [open/close]

          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. Biotechniques, 58 (2): 86-88, 2015) 。
          提供依頼 手続きの詳細は、「提供申込みについて[link]」をご覧ください。営利目的利用についてはお問い合わせください。
          提供依頼書 [Word]
          提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]

          Catalog # Resource name Shipping form Fee (non-profit org.)
          RDB18372 pUC18K_mPAG2 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

          Original reference

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

          Featured content

          Featured content Empty Backbone (English text)
          Featured content Empty Backbone (Japanese text)

          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 RDB18372_B0H5p1-1.pdf check

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

          Primer: KanR-C_F
          Region: KanR 3', mPAG2 5'
          Sequence file: RDB18372_B0H5a.seq check
          >D07025A1_B0H5_3_KanR-C_F_A01_01_ABI08.ab1
              1 NNNNNNNNNN NNNNNNAANN NNGNTNNTGA NAATCCTGNT ATGAATAAAT TGCAGTTTCA
             61 TTTGATGCTC GATGAGTTTT TCTAAAGGAG GTAAAACACA TGTCACACCT AGCAGAATTA
            121 GTTGCTAGTG CAAAAGCTGC AATATCACAA GCTAGCGATG TTGCGGCGCT GGACAATGTG
            181 CGTGTGGAGT ACCTGGGTAA GAAAGGCCAC CTGACCCTGC AGATGACCAC CCTGCGTGAA
            241 CTGCCGCCGG AGGAACGTCC GGCGGCGGGT GCGGTTATCA ACGAGGCGAA GGAACAAGTG
            301 CAGCAAGCGC TGAACGCGCG TAAAGCGGAG CTGGAAAGCG CGGCGCTGAA CGCGCGTCTG
            361 GCGGCGGAGA CCATTGACGT TAGCCTGCCG GGTCGTCGTA TTGAAAACGG TGGCCTGCAC
            421 CCGGTGACCC GTACCATCGA TCGTATTGAG AGCTTCTTTG GTGAACTGGG CTTCACCGTT
            481 GCGACCGGTC CGGAGATCGA AGACGATTAT CACAACTTTG ATGCGCTGAA CATTCCGGGT
            541 CACCACCCGG CGCGTGCGGA CCACGATACC TTCTGGTTTG ATACCACCCG TCTGCTGCGT
            601 ACCCAGACCA GCGGTGTTCA AATCCGTACC ATGAAGGCGC AGCAACCGCC GATTCGTATC
            661 ATTGCGCCGG GCCGTGTGTA CCGTAACGAC TATGATCAGA CCCACACCCC GATGTTCCAC
            721 CAAATGGAGG GTCTGATCGT GGACACCAAC ATTAGCTTCA CCAACCTGAA AGGCACCCTG
            781 CACGACTTTC TGCGTAACTT CTTTGANGAA GATCTGCAGA TCCGTTTTCG TCCGANCTAC
            841 TTCCCGTTTG CGGANCCNAN CGCGGAAGTG NANGTTATGG GCAANAACGG CAAATGGCTG
            901 GANNTTCTGG GTTGCGGNAT GGNGCACCNG ANCGTTCTGC NNAACGTGGG NATNGNNNCN
            961 GNANNNGTAC AGNNGNTTTC NGCTTNNGNT ATNGGNNNNG NNNNGNNNGA NNNNNCNNNN
          1021 NNNNNN
          //
          Primer: ePAG_R1
          Region: mPAG2 3'
          Sequence file: RDB18372_B0H5b.seq check
          >D07025A1_B0H5_ePAG_R1_B01_02_ABI08.ab1
              1 NNNNNNNNNN NNNNNNAANN GNNCTTNNNC TAGNTCCTTT TAAATTAAAA ATGAAGTTTT
             61 AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GGAATTATTA CTTAAATTGT
            121 TTCAGGAAGC GCAGATCGTT TTCAAAGAAG CTACGCAGGT CGGTCACGCC ATAACGCAGC
            181 ATGGTCAGAC GTTCCATGCC CATACCAAAG CCGAAACCGC TGTACACTTC CGGATCAATA
            241 CCCACGTTAC GCAGAACGTT CGGGTGCACC ATGCCGCAAC CCAGAACTTC CAGCCATTTG
            301 CCGTTCTTGC CCATAACGTC CACTTCCGCG CTCGGCTCCG CAAACGGGAA GTAGCTCGGA
            361 CGAAAACGGA TCTGCAGATC TTCCTCAAAG AAGTTACGCA GAAAGTCGTG CAGGGTGCCT
            421 TTCAGGTTGG TGAAGCTAAT GTTGGTGTCC ACGATCAGAC CCTCCATTTG GTGGAACATC
            481 GGGGTGTGGG TCTGATCATA GTCGTTACGG TACACACGGC CCGGCGCAAT GATACGAATC
            541 GGCGGTTGCT GCGCCTTCAT GGTACGGATT TGAACACCGC TGGTCTGGGT ACGCAGCAGA
            601 CGGGTGGTAT CAAACCAGAA GGTATCGTGG TCCGCACGCG CCGGGTGGTG ACCCGGAATG
            661 TTCAGCGCAT CAAAGTTGTG ATAATCGTCT TCGATCTCCG GACCGGTCGC AACGGTGAAG
            721 CCCAGTTCAC CAAAGAAGCT CTCAATACGA TCGATGGTAC GGGTCACCGG GTGCAGGCCA
            781 CCGTTTTCNA TACGACGANC CGGCANGCTA ACGTCNATGG TCTCCGCCGC CAGANGCGCG
            841 TTCAGCGCCG CGCTTTCCAG CTNCGCTTTA CGCGCGTTCA GCGCTNGCTG CACTTGTNCC
            901 TTCGCCNCGT TGANAANCGC ACCNGCCGCC NGANGTTCCN NNGGCGGCAG TTCNCGCNNG
            961 GNGGNCATCT GCNNGNNNNN NTGGNNNTTT NNNANCNNNG NNANTNNNNA CNNNCATTGN
          1021 CCNNNNNNNN NN
          //
          Primer: pGEX3'
          Region: AmpR pro, KanR5'
          Sequence file: RDB18372_B0H5c.seq check
          >D07025A1_B0H5_3_pGEX3'_E02_05_ABI08.ab1
              1 NNNNNNNNNN NNNCGCNNNN NNNNNNGNCC TCGTGNNACG CCTATTTTTA TAGGTTAATG
             61 TCATGATAAT AATGGTTTCT TAGACGTCAG GTGGCACTTT TCGGGGAAAT GTGCGCGGAA
            121 CCCCTATTTG TTTATTTTTC TAAATACATT CAAATATGTA TCCGCTCATG AGACAATAAC
            181 CCTGATAAAT GCTTCAATAA TATTGAAAAA GGAAGAGTAT GAGCCATATT CAACGGGAAA
            241 CGTCTTGCTC GAGGCCGCGA TTAAATTCCA ACATGGATGC TGATTTATAT GGGTATAAAT
            301 GGGCTCGCGA TAATGTCGGG CAATCAGGTG CGACAATCTA TCGATTGTAT GGGAAGCCCG
            361 ATGCGCCAGA GTTGTTTCTG AAACATGGCA AAGGTAGCGT TGCCAATGAT GTTACAGATG
            421 AGATGGTCAG ACTAAACTGG CTGACGGAAT TTATGCCTCT TCCGACCATC AAGCATTTTA
            481 TCCGTACTCC TGATGATGCA TGGTTACTCA CCACTGCGAT CCCCGGGAAA ACAGCATTCC
            541 AGGTATTAGA AGAATATCCT GATTCAGGTG AAAATATTGT TGATGCGCTG GCAGTGTTCC
            601 TGCGCCGGTT GCATTCGATT CCTGTTTGTA ATTGTCCTTT TAACAGCGAT CGCGTATTTC
            661 GTCTCGCTCA GGCGCAATCA CGAATGAATA ACGGTTTGGT TGATGCGAGT GATTTTGATG
            721 ACGAGCGTAA TGGCTGGCCT GTTGAACAAG TCTGGAAAGA AATGCATAAA CTTTTGCCAT
            781 TCTCACCGGA TTCAGTCGTC ACTCATGGTG ATTTCTCACT TGATAACCTT ATTTTTGACG
            841 AGGGGAAATT AATANGTTGT ATTGATGTTG GANGANNCGG ANTCGCANAN CGATACCANG
            901 ATCTTGCCNT NCTATGGANN TGNCNCNGTG AGTTTTCTCC TTCNTTACNN AAACNGCTTT
            961 TTCNAAAATA TGNNANTGAN NATCNNGANN NGNNNAAANN GNANNTTCNT TTNNATGNNC
          1021 NNNNANNTTT TNNNNANNNN NNNNAANNNN NNNNNN
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

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

          2022.08.12

          GNP_filter3_RDBDEP_html_220812.pl