LOCUS       sequence-458353-        4618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mutant E. coli TrpRSTyrRS from a glnS promoters for 
            proteome-wide incorporation of tryptophan/tyrosine analogues..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4618)
  AUTHORS   Loynd C, Singha Roy SJ, Canarelli SE, Bak DW, Sundaresh B, Babbitz 
            Z, van Opijnen T, Weerapana E, Chatterjee A
  TITLE     Cell-selective multiplexed bioorthogonal noncanonical amino acid 
            tagging for nascent proteomics.
  JOURNAL   Nat Chem Biol. 2025 Oct 2. doi: 10.1038/s41589-025-02039-3.
  PUBMED    41039161
REFERENCE   2  (bases 1 to 4618)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2025 Oct 2. doi: 10.1038/s41589-025-02039-3."
COMMENT     Sequence Label: pBK-EcTrpRS(H14)-EcTyrRS(VSMA,D165G)
FEATURES             Location/Qualifiers
     source          1..4618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(239..827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(319..338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(938..957)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             949..1764
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             949..1764
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1022..1041)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tgcctgatgc gctacgctta tcaggcctac atgatctctg caatatattg agtttgcgtg
       61 cttttgtagg ccggataagg cgttcacgcc gcatccggca agaaacagca aacaatccaa
      121 aacgccgcgt tcagcggcgt tttttctgct tttcttcgcg aattaattcc gcttcgcaca
      181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      661 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
      721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
      841 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      901 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
      961 caacgggaaa cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1021 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     1081 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1141 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1201 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa
     1261 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     1321 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     1381 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     1441 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag
     1501 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     1561 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     1621 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     1681 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     1741 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     1801 attacgctga cttgacggga cggcggcttt gttgaataaa tcgaactttt gctgagttga
     1861 aggatccgcg gccgcgtgcg aagcggaatt aattcgcgaa gaaaagcaga aaaaacgccg
     1921 ctgaacgcgg cgacccgagc ggccgctcgg gttgtcagcc tgtcccgctt ataagatcat
     1981 acgccgttat acgttgttta cgctttgagg aatcccatat ggcaagcagt aacttgatta
     2041 aacaattgca agagcggggg ctggtagccc aggtgacgga cgaggaagcg ttagcagagc
     2101 gactggcgca aggcccgatc gcgctcgtgt gcggcttcga tcctaccgct gacagcttgc
     2161 atttggggca tcttgttcca ttgttatgcc tgaaacgctt ccagcaggcg ggccacaagc
     2221 cggttgcgct ggtaggcggc gcgacgggtc tgattggcga cccgagcttc aaagctgccg
     2281 agcgtaagct gaacaccgaa gaaactgttc aggagtgggt ggacaaaatc cgtaagcagg
     2341 ttgccccgtt cctcgatttc gactgtggag aaaactctgc tatcgcggcg aacaactatg
     2401 actggttcgg caatatgaat gtgctgacct tcctgcgcga tattggcaaa cacttctccg
     2461 ttaaccagat gatcaacaaa gaagcggtta agcagcgtct caaccgtgaa ggtcagggga
     2521 tttcgttcac tgagttttcc tacaacctgt tgcagggtta tagtatggcc tgtgcgaaca
     2581 aacagtacgg tgtggtgctg caaattggtg gttctgacca gtggggtaac atcacttctg
     2641 gtatcgacct gacccgtcgt ctgcatcaga atcaggtgtt tggcctgacc gttccgctga
     2701 tcactaaagc agatggcacc aaatttggta aaactgaagg cggcgcagtc tggttagatc
     2761 cgaagaaaac cagcccgtac aaattctacc agttctggat caacactgcg cgtgccgacg
     2821 tttaccgctt cctgaagttc ttcaccttta tgagcattga agagatcaac gccctggaag
     2881 aagaagataa aaacagcggt aaagcaccgc gcgcccagta tgtactggcg gagcaggtga
     2941 ctcgtctggt tcacggtgaa gaaggtttac aggcggcaaa acgtattacc gaatgcctgt
     3001 tcagcggttc tttgagtgcg ctgagtgaag cggacttcga acagctggcg caggacggcg
     3061 taccgatggt tgagatggaa aagggcgcag acctgatgca ggcactggtc gattctgaac
     3121 tgcaaccttc ccgtggtcag gcacgtaaaa ctatcgcctc caatgccatc accattaacg
     3181 gtgaaaaaca gtccgatcct gaatacttct ttaaagaaga agatcgtctg tttggtcgtt
     3241 ttaccttact gcgtcgcggt aaaaagaatt actgtctgat ttgctggaaa taaccatggc
     3301 tgcagtttca aacgctaaat tgcctgatgc gctacgctta tcaggcctac atgatctctg
     3361 caatatattg agtttgcgtg cttttgtagg ccggataagg cgttcacgcc gcatccggca
     3421 agaaacagca aacaatccaa aacgccgcgt tcagcggcgt tttttctgct tttcttcgcg
     3481 aattaattcc gcttcgcacg cggccgctcg ggttgtcagc ctgtcccgct tataagatca
     3541 tacgccgtta tacgttgttt acgctttgag gaatcccata tgatgactaa gcccatcgtt
     3601 tttgctggcg cacagccctc aggtgaattg accattggta actacatggg tgcgctgcgt
     3661 cagtgggtaa acatgcagga tgactaccat tgcatttact gtatcgttga ccaacacgcg
     3721 atcaccgtgc gccaggatgc acagaagctg cgtaaagcga cgctggatac gctggccttg
     3781 tatctggctt gtggtatcga tcctgagaaa agcaccattt ttgttcagtc ccacgtgccg
     3841 gaacatgcac agttaggctg ggcactgaac tgctatacct acttcggcga actgagtcgc
     3901 atgacgcagt ttaaagataa atctgcgcgt tatgccgaga acatcaacgc tggtctgttt
     3961 gactatccgg tgctgatggc agcggacatc ctgctgtatc aaactaatct gggtccttgt
     4021 ggtgaagacc agaaacagca cctcgaactg agccgcgata ttgcccagcg tttcaacgcg
     4081 ctgtatggcg agatctttaa ggtgccggag ccgtttattc cgaaatctgg cgcgcgcgta
     4141 atgtcgctgc tggagccgac caagaagatg tccaagtctg acgataatcg caataacgtt
     4201 atcggcctgc tggaagatcc gaaatcggta gtgaagaaaa tcaaacgtgc ggtcactgac
     4261 tccgacgagc cgccggtagt tcgctacgat gtgcagaaca aagcgggcgt ttccaacctg
     4321 ttggatatcc tttcagcggt aacgggccag agcatcccag aactggaaaa acagttcgaa
     4381 ggcaagatgt atggtcatct gaaaggtgaa gtggctgatg ccgtttccgg tatgctgact
     4441 gaattgcagg aacgctatca ccgtttccgc aacgatgaag ccttcctgca acaggtgatg
     4501 aaagatggcg cggaaaaagc cagcgcgcac gcttcccgta cgctaaaagc ggtgtacgaa
     4561 gcgattggtt ttgtggcgaa gccgtaaggt accatggctg cagtttcaaa cgctaaat
//
