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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.
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 |
 
Other clones in our bank Escherichia coli (KEGG orthology K01889) | E. coli (KEGG orthology K19272) |

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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RDB17947 pUC18K_ePAG2 DNA solution

Ordering Information [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

Electronic file Nucleotide sequence provided by the depositor RDB17947zzk01.seq

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.

2021.01.06

GNP_filter3_RDBDEP_html_201222.pl